WO1996013575A1

WO1996013575A1 - Recombinant feline herpes virus

Info

Publication number: WO1996013575A1
Application number: PCT/US1995/013975
Authority: WO
Inventors: Mark D. Cochran; Michael W. Mcdonnell
Original assignee: Syntro Corporation
Priority date: 1994-10-26
Filing date: 1995-10-26
Publication date: 1996-05-09
Also published as: AU4197196A

Abstract

The present invention provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains a deletion in the unique short region of the viral genome, wherein the deletion is in the glycoprotein E (gE) gene.

Description

RECOMBINANT FELINE HERPES VIRUS

This application is a continuation of United States Application Serial No. 08/329,883, filed on October 26, 1994, the contents of which are hereby incorporated by reference.

Throughout this application various publications are referenced by Arabic numerals in brackets. Pull citations for these publications may be found at the end of the specification immediately preceding the claims. The disclosures of these publications are in their entirety hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains.

BACKGROUND OF THE INVENTION

Herpesviruses contain 100,000 to 200,000 base pairs of DNA as their genetic material having a long unique segment and a short unique segment, the short unique segment bounded by an internal repeat sequence and a terminal repeat sequence. Within a given genome, several regions have been identified that are not essential for the replication of virus in cell culture. Modifications of some of these^' regions of the DNA have been known to lower the pathogenicity of the virus, i.e., to attenuate the virus when it infects an animal species. For example, inactivation of the thymidine kinase gene of either human herpes simplex virus [1] or pseudorabies virus of swine [2] renders the human herpesvirus less pathogenic in mice and the pseudorabies virus less pathogenic in swine.

Removal of specific regions of the repeat region of a human herpes simplex virus has been shown to render the virus less pathogenic [1, 3] . Furthermore, a repeat region has been identified in Marek's disease virus that is associated with viral oncogenicity [4] . A region in herpesvirus saimiri has similarly been correlated with oncogenicity [5] . Removal of a specific region of the repeat region renders pseudorabies virus less pathogenic (US Pat. 4,877,737) . A region in pseudorabies virus has been shown to be deleted in naturally-occurring vaccine strains [6] . These deletions are at least in part responsible for the lack of pathogenicity of these strains. Specific combinations of glycoproteins gene deletions in herpes simplex virus render the herpes simplex virus less pathogenic in mice [see 37 for review] . While, combinations of glycoprotein gene deletions in pseudorabies virus render the pseudorabies virus less pathogenic in swine [see 38 for review] .

Herpesviruses contain non-essential regions of DNA in various parts of the genome, and that modification of these regions can attenuate the virus, leading to a non-pathogenic strain from which a vaccine may be derived. The degree of attenuation of the virus is important to the utility of the virus as a vaccine. Deletions which cause too much attenuation of the virus will result in a vaccine that fails to elicit an adequate immune response. Although several examples of attenuating deletions are known, the appropriate combination of deletions for any herpesvirus is not readily apparent.

Feline herpesvirus 1 (FHV) is the causative agent of feline viral rhinotracheitis, an acute upper respiratory disease in cats [7, 8] . Serological studies indicate that 50 to 70% of adult cats have detectable antibodies to the virus [9, 10] . Currently available inactivated and attenuated live virus vaccines reduce disease but do not prevent infection by FHV [11] .

The feline herpesvirus is a member of the family herpesviridae, which are commonly known as the herpesviruses and a member of the subfamily alphaherpesvirus with a group

D genome [12] . The FHV genome is comprised of approximately 134 kilobase (kb) pairs that is subdivided into a long unique segment of approximately 104 kb and a short unique segment of approximately 8 kb [13] . The unique short region is bounded by inverted repeat sequences which are approximately 11 kb. Physical maps of restriction endonuclease sites of the FHV genome have been published

[12, 13] . The thymidine kinase gene of FHV has been sequenced and an FHV virus containing a deletion of the TK gene was isolated using a drug selection technique [14] . An FHV having a deletion of the TK gene and an insertion of the feline leukemia virus (FeLV) envelope (eπv) and gag genes at the TK deletion site has been constructed [36] .

The feline herpesviruses in this invention are useful as vectors for the delivery of vaccine antigens from microorganisms causing diseases in animals other than cats or dogs and for the delivery of therapeutic agents. The therapeutic agent that is delivered by a viral vector of the present invention must be a biological molecule that is a by-product of feline herpesvirus replication. This limits the therapeutic agent in the first analysis to either DNA, RNA or protein. There are examples of therapeutic agents from each of these classes of compounds in the form of anti- sense DNA, anti-sense RNA [18] , ribozy es [19] , suppressor tRNAs [20] , interferon-inducing double stranded RNA and numerous examples of protein therapeutics, from hormones, e.g., insulin, to lymphokines, e.g., interferons and interleukins, to natural opiates. The discovery of these therapeutic agents and the elucidation of their structure and function does not necessarily allow one to use them in a viral vector delivery system. SUMMARY OF THE INVENTION

This invention provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains a deletion in the unique short region of the viral genome, wherein the deletion is in the glycoprotein E (gE) gene.

This invention provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains an insertion into the unique short region of the viral genome, wherein the insertion is in the glycoprotein E (gE) gene. Homology vectors for producing a recombinant feline herpes virus and host cells are provided for.

This invention provides a vaccine for feline herpes virus which comprises an effective immunizing amount of the recombinant feline herpes virus and a suitable carrier. A method of immunizing an animal against a human or feline pathogen is also provided for.

This invention provides a method of distinguishing an animal vaccinated with the feline herpes virus vaccine from an animal infected with a naturally-occuring feline herpes virus.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1: Restriction endonuclease map of the feline herpesvirus genome.

Figure 2: Feline leukemia virus envelope epitopes

1 and 3 (FeLV EE #1 and FeLV EE #3; SEQ

ID NOs: 7, 8, 9, 10) . DNA and amino acid sequences of the antigenic epitopes of the FeLV envelope gene

[21] . DNA sequences have compatible

BamΑI and Bgrlll ends so that multiple copies of each epitope could be cloned as fusion proteins in the FHV homology vectors.

Figures 3A-3D: DNA insertion in Homology Vectors 411-

91.01A and 411-91.01B. The diagram shows the orientation of DNA fragments assembled in plasmid 411-91.01A and

411-91.01B. The source of each fragment is described in the Materials and Methods section. The source of FHV DNA sequences is FHV homology vector 416-80.21B. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 11) , junction B (SEQ ID NO: 12) , junction C (SEQ ID NO: 13) , junction D (SEQ ID NO: 14) and junction E (SEQ ID

NO: 15) . The restriction endonuclease sites used to generate each fragment as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , human cytomegalovirus (HCMV) , immediate early

(IE) , feline leukemia virus (FeLV) , pseudorabies virus (PRV) , polyadenylation site (poly A) .

Figures 4A-4B: DNA insertion in Homology Vector 416-

80.2IB. The diagram shows the orientation of DNA fragments assembled in plasmid 416-80.21B. The source of each fragment is described in the Materials and Methods section. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 16) , junction B (SEQ ID NO: 17) , and junction C (SEQ ID NO: 18) . The restriction endonuclease sites used to generate each fragment ,as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The location of the gl gene coding regions is also given. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , glycoprotein I (gl) and glycoprotein E (gE) .

Figures 5A-5C: DNA insertion in Homology Vectors 416-

88.2L. The diagram shows the orientation of DNA fragments assembled in plasmid 416-88.2L. The source of each fragment is described in the Materials and Methods section. The origin of FHV DNA sequences is FHV homology vector 416-80.2IB. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 19) , junction B (SEQ ID NO: 20) , junction C (SEQ ID NO: 21), and junction D (SEQ ID

NO: 22) . The restriction endonuclease sites used to generate each fragment as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , pseudorabies virus (PRV) , glycoprotein X (gX) , polyadenylation site (poly A) .

Figure 6 : DNA insertion at the gE deletion site of S-FHV-010. The diagram shows the synthetic DNA inserted at the gE deletion site and the unique Sfil restriction endonuclease site of the recombinant S-FHV-010. The sequences - 8 - located at the junctions between each fragment are shown (SEQ ID NO: 23) . The restriction endonuclease sites used to generate each fragment as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviation is used, feline herpesvirus (FHV) .

Figures 7A-7E: DNA insertion in Homology Vectors 478-

17.IT and 478-10.11. The diagram shows the orientation of DNA fragments assembled in plasmid 478-17. IT and 478-

10.11. The source of each fragment is described in the Materials and Methods section. The origin of FHV DNA sequences is FHV homology vector 416- 80.21B. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 24) , junction B (SEQ ID NO: 25) , junction C (SEQ ID NO: 26) , junction D (SEQ ID NO: 27), junction E (SEQ ID NO:

28), junction F (SEQ ID NO: 29) , junction G (SEQ ID NO: 30) , junction H (SEQ ID NO: 31), junction I (SEQ ID NO: 32) , and junction J (SEQ ID NO: 33) . The restriction endonuclease sites used to generate each fragment as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , human cytomegalovirus (HCMV) , immediate early

(IE) , feline leukemia virus (FeLV) , pseudorabies virus (PRV) , glycoprotein

X (gX) , Escherichia coli (E. coli ) , herpes simplex virus type 1 (HSV-1) , polyadenylation site (poly A) .

Figures 8A-8E: DNA insertion in Homology Vectors 510-

36.IC and 510-36.2F. The diagram shows the orientation of DNA fragments assembled in plasmid 510-36. IC and 510- 36.2F. The source of each fragment is described in the Materials and Methods section. The origin . of FHV DNA sequences is FHV homology vector 416-

80.21B. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 34) , junction B (SEQ ID NO: 35), junction C (SEQ ID NO: 36) , junction D

(SEQ ID NO: 37), junction E (SEQ ID NO: 38) , junction F (SEQ ID NO: 39) , junction G (SEQ ID NO: 40) , junction H (SEQ ID NO: 41) , and junction I (SEQ ID NO: 42) . The restriction endonuclease sites used to generate each fragment as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , human cytomegalovirus (HCMV) , immediate early

(IE) , feline leukemia virus (FeLV) , pseudorabies virus (PRV) , glycoprotein

Figures 9A-9E: DNA insertion in Homology Vectors 639-

85.IH. The diagram shows the orientation of DNA fragments assembled in plasmid 639-85.1H. The source of each fragment is described in the Materials and Methods section. The origin of FHV DNA sequences is FHV homology vector 644-09.B2. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 43) , junction B (SEQ ID NO: 44) , junction C (SEQ ID NO: 45) , junction D (SEQ ID NO:

46) , junction E (SEQ ID NO: 47) , junction F (SEQ ID NO: 48) and junction G _(SEQ ID NO: 49) . The restriction endonuclease sites used to generate each fragment as well as the synthetic

DNA sequences that were used to join - the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , human cytomegalovirus (HCMV) , immediate early

(IE) , Dirofilaria immi tis (D. immi tis) , pseudorabies virus (PRV) , glycoprotein X (gX) , Escherichia coli (E. coli ) , herpes simplex virus type 1 (HSV-1) , polyadenylation site (poly A) .

Figures 10A-10C:

DNA insertion in Homology Vectors 644- 09.A1. The diagram shows the orientation of DNA fragments assembled in plasmid 644-09.Al. The source of each fragment is described in the Materials and Methods section. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 50) , junction B (SEQ ID NO: 51) and junction C (SEQ ID NO: 52) . The restriction endonuclease sites used to generate each fragment as well as the synthetic

DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during cons t ru c t ion . The f o l l owi ng abbreviations are used, feline herpesvirus (FHV) , glycoprotein I (gl ) and glycoprotein E (gE) .

Figures 11A-11B :

DNA insertion in Homology Vectors 644- 09.B2. The diagram shows the orientation of DNA fragments assembled in plasmid 644-09.B2. The source of each fragment is described in the Materials and Methods section. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 53) , junction B (SEQ ID NO: 54) and junction C (SEQ ID NO: 55) . The restriction endonuclease sites used to generate each fragment as well as the synthetic

DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following abbreviations are used, feline herpesvirus (FHV) , glycoprotein I (gl) and glycoprotein E (gE) .

Figures 12A-12E: DNA insertion in Homology Vectors 669-

42.04. The diagram shows the orientation of DNA fragments assembled in plasmid 669-42.04. The source of each fragment is described in the Materials and Methods section. The origin of FHV DNA sequences is FHV homology vector 644-09.B2. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 56) , junction B (SEQ ID NO: 57) , junction C (SEQ ID NO: 58), junction D (SEQ ID NO:

59) , junction E (SEQ ID NO: 60) , junction F (SEQ ID NO: 61) , junction G

(SEQ ID NO: 62) and junction H (SEQ ID

NO: 63) . The restriction endonuclease sites used to generate each fragment as well as the synthetic DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during- construction. The following abbreviations are used, feline herpesvirus (FHV) , human cytomegalovirus (HCMV) , immediate early (IE) , feline immunodeficiency virus (FIV) , pseudorabies virus (PRV) , glycoprotein X (gX) , Escherichia coli

(E. coli) , herpes simplex virus type 1 (HSV-1) , polyadenylation site (poly A) .

Figures 13A-13E: DNA insertion in Homology Vectors 725-

26.A10. The diagram shows the orientation of DNA fragments assembled in plasmid 725-26.A10. The source of each fragment is described in the Materials and Methods section. The origin of FHV DNA sequences is FHV 5 homology vector 644-09.B2. The sequences located at the junctions between each fragment are shown, including junction A (SEQ ID NO: 64) , junction B (SEQ ID NO: 65) , junction C

10 (SEQ ID NO: 66), junction D (SEQ ID NO:

67) , junction E (SEQ ID NO: 68), junction F (SEQ ID NO: 69) and junction G (SEQ ID NO: 70) . The restriction endonuclease sites used to generate

15 each fragment as well as the synthetic

DNA sequences that were used to join the fragments are described for each junction. The synthetic DNA sequences are underlined by a solid bar. The

20 following convention is used: restriction endonuclease sites in brackets [] indicate the remnants of sites that were destroyed during construction. The following

25 abbreviations are used, feline herpesvirus (FHV) , human cytomegalovirus (HCMV) , immediate early (IE) , feline immunodeficiency virus (FIV) , pseudorabies virus (PRV) ,

30 glycoprotein X (gX) , Escherichia coli

(E. coli) , herpes simplex virus type 1 (HSV-1) , thymidine kinase (TK) , polyadenylation site (poly A) .

35 DETAILED DESCRIPTION OF THE INVENTION

The present invention involves recombinant feline herpesviruses useful in the preparation of vaccines to protect cats from naturally-occuring infectious feline herpesvirus. The present invention is also useful for expression of foreign genes from other pathogens for protection against disease. Recombinant feline herpesvirus expressing foreign genes from avian or mammalian pathogens are useful as vaccines in avian or mammalian species including but not limited to chickens, turkeys, ducks, feline, canine, bovine, equine, and primate, including human.

The present invention provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains a deletion in the unique short region of the viral genome, wherein the deletion is in the glycoprotein E (gE) gene. Said recombinant feline herpes virus contains a deletion which attenuates the virus, rendering it suitable for use as a vaccine against feline herpesvirus.

In one embodiment the feline herpes virus contains a foreign DNA sequence inserted into a non-essential region of the feline herpes virus genome. In another embodiment the foreign DNA sequence is inserted into a unique short region of the feline herpes virus. In another embodiment the foreign DNA sequence is inserted in the deleted gE gene.

In a preferred embodiment the recombinant feline herpes virus is designated S-FHV-004. S-FHV-004 was deposited on October 26, 1994 under ATCC Accession No. VR 2487 pursuant to the Budapest Treaty on the International Deposit of Microorganisms for the Purposes of Patent Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.S.A.

In another preferred embodiment the recombinant feline herpes virus is designated S-FHV-014. The S-FHV-014 virus was deposited on October 26, 1994 under ATCC Accession No. VR 2488 pursuant to the Budapest Treaty on the International Deposit of Microorganisms for the Purposes of Patent Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.S.A.

For purposes of this invention, "a recombinant feline herpes virus" is a live feline herpes virus which has been generated by the recombinant methods well known to those of skill in the art, e.g., the methods set forth in DNA TRANSFECTION FOR GENERATING RECOMBINANT VIRUS in Materials and Methods, and the virus has not had genetic material essential for the replication of the feline herpes virus deleted.

The present invention also provides viruses in which (a) DNA corresponding to the gG or gl genes have been deleted, and (b) DNA encoding gG or gl have been altered or deleted, and a foreign DNA sequence from a pathogen of an animal has been inserted. The present invention also provides viruses in which the gE gene is deleted in combination with the gG and/or gl genes.

In addition, a recombinant feline herpes virus which further comprises the foreign DNA sequence inserted into the unique short region, wherein the foreign DNA sequence is capable of being expressed in a recombinant feline herpes virus host cell. The foreign DNA sequence is inserted into the unique short region or any non-essential region of the feline herpes virus viral genome in such a way that it is capable of being expressed in a recombinant feline herpes virus infected host cell.

For purposes of this invention, "a non-essential site" of the feline herpes virus viral genome is a region of the viral genome which is not necessary for viral infection and replication.

The following non-essential sites of the feline herpes virus viral genome are preferred sites for inserting a foreign DNA sequence or gene into the virus, include but are not limited to: the glycoprotein E (gE) gene, the glycoprotein G (gG) gene, or the glycoprotein I (gl) gene. Other non-essential sites are known to those skilled in the art.

In addition, the foreign DNA sequence, which is inserted into a non-essential site in the feline herpes virus viral genome, encodes a screenable marker, such as E. coli B-galactosidase or E. coli beta- glucuronidase.

For purposes of this invention, a "polypeptide which is a detectable marker" includes but is not limited to: the bimer, trimer and tetramer form of the polypeptide. E. coli beta-galactosidase is a tetramer composed of four polypeptides or monomer sub-units.

In addition, the foreign DNA sequence which is inserted into a non-essential site in the feline herpes virus viral genome, when introduced into the host cell, induces production of protective antibodies against a feline disease causing agent from which the antigen is derived or derivable. Such antigenic polypeptide may be derived or derivable from the following: feline pathogen, canine pathogen, equine pathogen, bovine pathogen, avian pathogen, porcine pathogen, or human pathogen.

Antigenic polypeptides include, but are not limited to: infectious bronchitis virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus, infectious laryngotracheitis virus, infectious bursal disease virus VP2 protein, infectious bursal disease virus VP3 protein, infectious bursal disease virus VP4 protein, Marek's disease virus glycoprotein gB, Marek's disease virus glycoprotein gA, Marek's disease virus glycoprotein gD, Newcastle disease virus fusion (F) protein, Newcastle disease virus hemagglutinin-neuraminidase (HN) , infectious laryngotracheitis virus glycoprotein I, infectious laryngotracheitis virus glycoprotein D, infectious laryngotracheitis virus glycoprotein B, infectious bronchitis virus spike protein, or infectious bronchitis virus matrix protein.

Such antigenic polypeptide may also be derived or derivable from avian encephalomyelitis virus, avian reovirus, avian paramyxovirus, avian influenza virus, avian adenovirus, fowl pox virus, avian coronavirus, avian rotavirus, chick anemia agent, Salmonella spp . , E. coli . , Pasteurella spp. , Bordetella spp. Eimeria spp. Histomonas spp. , Trichomonas spp. , poultry nematodes, cestodes, trematodes, poultry mites/lice, poultry protozoa.

The foreign DNA sequence or gene may be put under control of an endogenous upstream feline herpes virus promoter, or it may be put under control of a heterologous upstream promoter. The heterologous upstream promoter may be derived from the HCMV IE promoter, the PRV gX promoter, or BHV-1.1 VP8 promoter. In another embodiment, the promoter is selected from a group consisting of MDV gB promoter, MDV gA promoter, MDV gD promoter, ILTV gB promoter, ILTV gD promoter, or HSV-1 alpha 4 promoter.

The foreign DNA sequence or genes include, but are not limited to: feline leukemic virus envelope gene, hepatitus B core antigen gene; Pseudorabies virus glycoprotein C gene; Dirofilaria immi tis 22 kD or p39 gene; feline immunodeficiency virus gag, pol, or env; or E. Coli beta-galactosidase gene . In one embodiment the foreign DNA seuquence is a fusion protein two or more foreign DNA sequences or genes.

The present invention further provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains a deletion or other alteration in the unique short region of the viral genome, wherein the deletion or alteration is in the glycoprotein gE gene, so that upon replication, the recombinant virus produces no glycoprotein gE. The following recombinant viruses are preferred embodiments of this invention: A recombinant feline herpes virus designated S-FHV-003, S-FHV-006, S-FHV-007, S-FHV-009, S-FHV-012, S-FHV-008, S-FHV-011, S-FHV-005, S-FHV-010, S-FHV-016, or S-FHV-017.

The present invention further provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains a deletion or other alteration in the unique short region of the viral genome, wherein the deletion or alteration is in the glycoprotein gl gene, so that upon replication, the recombinant virus produces no glycoprotein gl.

The present invention further provides a recombinant feline herpes virus comprising the feline herpes virus viral genome which contains a deletion or other alteration in the unique short region of the viral genome, wherein the deletion or alteration is in the glycoprotein gG gene and in the glycoprotein gl gene, so that upon replication, the recombinant virus produces no glycoprotein gG and no glycoprotein gl .

It is contemplated that a deletion in any one the above genes will attenuate the virus, rendering it suitable to be used as a vaccine against feline herpesvirus. A foreign DNA sequence may be inserted within any one of these sites in such a way that it may be expressed in a host cell which is infected which the recombinant feline herpes virus of the present invention.

In one embodiment the recombinant feline herpes virus further contains a deletion in a unique short region of the feline herpes virus genome. In another embodiment the recombinant feline herpes virus is further characterized by a deletion in a glycoprotein I (gl) gene. In another embodiment the recombinant feline herpes virus is further characterized by a deletion in a glycoprotein G (gG) gene.

The present invention further provides a homology vector for producing a recombinant feline herpes virus by inserting a foreign DNA sequence into the feline herpes virus genome which comprises a double-stranded DNA molecule consisting of: a) double-stranded foreign DNA sequence encoding an antigenic polypeptide derived from a feline pathogen; b) at one end of the foreign DNA sequence, double-stranded feline virus genomic DNA homologous to the genomic DNA located at one side of a non-essential site of the feline herpes viral genomic DNA; c) at the other end of the foreign DNA sequence, double stranded feline herpes virus genomic DNA homologous to the genomic DNA located at the other side of the same site.

In another embodiment, the antigenic polypeptide of a human pathogen is derived from human herpesvirus, herpes simplex virus-1, herpes simplex virus-2, human cytomegalovirus, Epstein-Barr virus, Varicell-Zoster virus, human herpesvirus-6, human herpesvirus-7, human influenza, human immunodeficiency virus, rabies virus, measles virus, hepatitis B virus and hepatitis C virus. Furthermore, the antigenic polypeptide of a human pathogen may be associated with malaria or malignant tumor from the group conisting of Plasmodium falciparum, Bordetella pertusis, and malignant tumor.

In another embodiment the double stranded foreign DNA sequence in the homology vector encodes an antigenic polypeptide derived from an equine pathogen. The antigenic polypeptide of an equine pathogen can derived from equine influenza virus or equine herpesvirus. Examples of such antigenic polypeptide are equine influenza virus type A/Alaska 91 neuraminidase, equine influenza virus type A/Prague 56 neuraminidase, equine influenza virus type A/Miami 63 neuraminidase, equine influenza virus type A/Kentucky 81 neuraminidaseequine herpesvirus type 1 glycoprotein B, and equine herpesvirus type 1 glycoprotein D.

In another embodiment the double stranded foreign DNA sequence of the homology vector encodes an antigenic polypeptide derived from bovine respiratory syncytial virus or bovine parainfluenza virus. The antigenic polypeptide of derived from bovine respiratory syncytial virus equine pathogen can derived from equine influenza virus is bovine respiratory syncytial virus attachment protein (BRSV G) , bovine respiratory syncytial virus fusion protein (BRSV F) , bovine respiratory syncytial virus nucleocapsid protein (BRSV N) , bovine parainfluenza virus type 3 fusion protein, and the bovine parainfluenza virus type 3 hemagglutinin neuraminidase.

In another embodiment the double stranded foreign DNA sequence in the homology vector encodes a cytokine capable of stimulating human immune response. For example, the cytokine may be, but is not limited to, interleukin-2, interleukin-6, interleukin-12 , interferons, granulocyte-macrophage colony stimulating factors, and interleukin receptors.

Preferred embodiments of this invention are the homology vectors designated Homology Vector 416-80.21B, 644-09.Al, 644-09.B2, 478-17.IT, 478.10.11, 411-91.01A, 411-91.01B, 510-36.IC, 510-36.2F, 416-88.2L, 639.85.1H, 669-42.04 or 725-26.A10.

The present invention further provides a homology vector for producing a recombinant feline herpes virus by inserting a foreign DNA sequence into the feline herpes virus genome which comprises a double-stranded DNA molecule consisting of: a) double-stranded foreign DNA sequence encoding an antigenic polypeptide derived from a cytokine capable of stimulating an immune response; b) at one end of the foreign DNA sequence, double-stranded feline herpes virus genomic DNA homologous to the genomic DNA located at one side of a non-essential site of the feline herpes virus genomic

DNA; c) at the other end of the foreign DNA sequence, double stranded feline virus genomic DNA homologous to the genomic DNA located at the other side of the same site.

The present invention further provides a host cell infected with the recombinant feline herpes virus. In one embodiment the host cell is a mammalian cell. Other host cells are known to those skilled in the art.

The present invention further provides a vaccine for feline herpes virus which comprises an effective immunizing amount of the recombinant feline herpes virus and a suitable carrier. In one embodiment the vaccine against an feline pathogen comprises an effective immunizing amount of the recombinant feline herpes virus and a suitable carrier. This vaccine may contain either inactivated or live recombinant virus.

Suitable carriers for the recombinant virus are well known to those skilled in the art and include but are not limited to proteins, sugars, etc. One example of such a suitable carrier is a physiologically balanced culture medium containing one or more stabilizing agents such as hydrolyzed proteins, lactose, etc. Preferably, the live vaccine is created by taking tissue culture fluids and adding stabilizing agents such as stabilizing, hydrolyzed proteins. Preferably, the inactivated vaccine uses tissue culture fluids directly after inactivation of the virus.

The present invention further provides a method of immunizing an animal against a human pathogen which comprises administering to the animal an effective immunizing dose of the feline herpes vaccine. In one embodiment the method of immunizing an animal against an feline pathogen comprises administering to the animal an effective immunizing dose of the feline herpes vaccine.

The present invention further provides a multivalent vaccine for feline herpes virus and for one or more of other feline diseases which comprises an effective immunizing amount of a recombinant virus comprising the feline herpes virus viral genome which contains a deletion in the unique short region, wherein the deletion is in the glycoprotein E gene, and an insertion of a foreign gene into a non-essential site of the viral genome.

The foreign DNA sequence encodes an antigenic polypeptide which induces host cell production of protective antibodies against a feline disease causing agent from which the antigen is derived or derivable.

The invention further provides foreign RNA which encodes a polypeptide. Preferably, the polypeptide is antigenic in the animal. Preferably, this antigenic polypeptide is a linear polymer of more than 6 amino acids linked by peptide bonds which stimulates the animal to produce antibodies.

The present invention further provides a vaccine which comprises a suitable carrier and an effective immunizing amount of a recombinant feline herpes virus comprising the feline herpesvirus viral genome which contains a deletion or other alteration in the unique short region of the viral genome, wherein the deletion or alteration is in the glycoprotein E gene so that upon replication, the recombinant virus produces no glycoprotein E.

For purposes of this invention, an "effective immunizing amount" of the recombinant feline herpes virus of the present invention is within the range of IO³ to ^L0 PFU/dose. In another embodiment the immunizing amount is IO⁵ to fO PFU/dose. In a preffered embodiment the immunizing amount is IO⁶ PFU/dose. The method comprises administering to the animal an effective immunizing dose of the vaccine of the present invention. The vaccine may be administered by any of the methods well known to those skilled in the art, for example, by intramuscular, subcutaneous, intraperitoneal or intravenous injection. Alternatively, the vaccine may be administered intranasally or orally.

The present invention also provides a method of immunizing an animal, wherein the animal is a feline, canine, ovine, bovine, caprine, swine or human. For purposes of this invention, this includes immunizing the animal against the virus or viruses which cause the disease or diseases feline herpesvirus.

The present invention further provides a method of distinguishing an animal vaccinated with a feline herpes virus from an animal infected with a naturally- occuring feline herpes virus which comprises analysing a sample of a body fluid from the animal for the presence of feline herpes virus gE and at least one other antigen normally expressed in an animal infected by a naturally-occuring feline herpes virus, determining whether the antigen and gE are present in the body fluid, the presence of the antigen and the absence of gE indicative of an animal vaccinated with the vaccine and not infected with a naturally-occuring feline herpes virus.

In one embodiment the presence of the antigen and of gE in the body fluid is determined by detecting in the body fluid antibodies specific for the antigen and for gE.

The present invention provides a method for testing a feline to determine whether the feline has been vaccinated with the vaccine of the present invention, particularly the embodiment which contains the recombinant feline herpes virus S-FHV-004 or S-FHV-014.

This invention is further illustrated in the Experimental Details section which follows. This section is set forth to aid in an understanding of the invention but is not intended to, and should not be construed to, limit in any way the invention as set forth in the claims which follow thereafter.

EXPERIMENTAL DETAILS:

Materials and Methods

PREPARATION OF FHV VIRUS STOCK SAMPLES:

S-FHV-000 was obtained from the ATCC (ATCC No. 636) and S-FHV-001 was obtained from the NVSL (NVSL Challange Virus Strain SGE, Lot KS) . FHV virus stock samples were prepared by infecting Crandell Feline Kidney (CRFK) cells at a multiplicity of infection of 1.0 PFU/cell in Dulbecco's Modified Eagle Medium (DMEM) containing 2 mM glutamine, 100 units/ml penicillin, 100 units/ml streptomycin (these components were obtained from Irvine Scientific or equivalent supplier, and hereafter are referred to as complete DME medium) plus 5% fetal bovine serum. After cytopathic effect was complete, the medium and cells were harvested, aliquoted and frozen at -70°C. The titers were approximately 1 x IO⁷ to 1 x 10^B PFU/ml.

PREPARATION OF HERPESVIRUS DNA:

A confluent monolayer of CRFK cells in a 25 cm² flask or 60 mm petri dish was infected with 100 ml of virus sample. After overnight incubation, or when the cells were showing 100% cytopathic effect, the cells were scraped into the medium. The cells and medium were centrifuged at 3000 rpm for 5 minutes in a clinical centrifuge. The medium was decanted, and the cell pellet was gently resuspended in 0.5 ml solution containing 0.5% NONIDET P-40^® (octyl phenol ethylene oxide condensate containing an average of 9 moles of ethylene oxide per molecule) (NP-40^®, purchased from Sigma Chemical Co., St. Louis, MO.) . The sample was incubated at room temperature for 10 minutes. Ten ml of a stock solution of RNase A (Sigma Chemical Co., St. Louis, MO.) were added (stock was 10 mg/ml, boiled for 10 minutes to inactivate DNAse) . The sample was centrifuged to pellet nuclei. The DNA pellet was removed with a pasteur pipette or wooden stick and discarded. The supernatant fluid was decanted into a 1.5 ml Eppendorf tube containing 25 ml of 20% sodium dodecyl sulfate (Sigma) and 25 ml proteinase-K (10 mg/ml; Boehringer Mannheim Biochemicals, Indianapolis, IN) . The sample was mixed and incubated at 37°C for 30- 60 minutes. An equal volume of water-saturated phenol was added and the sample was mixed briefly. The sample was centrifuged in an Eppendorf minifuge for 5 minutes at full speed. The upper aqueous phase was removed to a new Eppendorf tube, and two volumes of absolute ethanol were added and the tube- put at -20°C for 30 minutes to precipitate nucleic acid. The sample was centrifuged in an Eppendorf minifuge for 5 minutes. The supernatant was decanted, and the pellet was air dried and rehydrated in -16 ml H₂0. For the preparation of larger amounts of DNA, the procedure was scaled up to start with roller bottles or 175 cm² flasks of CRFK cells. The DNA was stored in 0.01 M tris pH 7.5, 1 mM EDTA at 4°C.

MOLECULAR BIOLOGICAL TECHNIQUES: Techniques for the manipulation of bacteria and DNA, including such procedures as digestion with restriction endonucleases, gel electrophoresis, extraction of DNA from gels, ligation, phosphorylation with kinase, treatment with phosphatase, growth of bacterial cultures, transformation of bacteria with DNA, and other molecular biological methods are described in [22 and 23] . The polymerase chain reaction (PCR) was used to introduce restriction endonuclease sites convenient for the manipulation of various DNAs [24] . In general, amplified fragments were less than 500 base pairs in size and critical regions of amplified fragments were confirmed by DNA sequencing. Except as noted, these techniques were used with minor variations.

LIGATION:

DNA was joined together by the action of the enzyme T4 DNA ligase (BRL) . Ligation reactions contained various amounts of DNA (from 0.2 to 20mg) , 20mM Tris pH 7.5, lOmM MgCl₂, lOmM dithiothreitol (DTT) , 200 mM ATP and 20 units T4 DNA ligase in 10-20 ml final reaction volume. The ligation proceeded for 3-16 hours at 15°C.

DNA SEQUENCING:

Sequencing was performed using the USB Sequenase Kit and ³⁵S-dATP (NEN) . Reactions using both the dGTP mixes and the dITP mixes were performed to clarify areas of compression. Alternatively, compressed areas were resolved on formamide gels. Templates were double- stranded plasmid subclones or single stranded M13 subclones, and primers were either made to the vector just outside the insert to be sequenced, or to previously obtained sequence. Alternatively, DNA sequencing was performed on the Applied Biosystems Automated Sequencer Model 388A per instructions of the manufacturer using Taq DNA polymerase and fluorescently-labelled dideoxynucleotides. The sequence obtained was assembled and compared using DNAStar software. Subsequent manipulation and comparison of sequences obtained was performed with Superclone and Supersee programs from Coral Software and DNAStar.

SOUTHERN BLOTTING OF DNA:

The general procedure for Southern blotting was performed as in Maniatis et al. and Sambrook et al. [22, 23] . DNA was blotted to nitrocellulose filters and hybridized to appropriately labeled DNA probes. Probes for Southern blots were prepared using either the Nonradioactive DNA Labeling and Detection Kit of Boehringer Mannheim or the nick translation kit of Bethesda Research Laboratories (BRL) . In both cases the manufacturer's recommended procedures were followed.

DNA TRANSFECTION FOR GENERATING RECOMBINANT VIRUS:

The method is based upon the calcium phosphate procedure of Graham and Van der eb [25] with the following modifications. Virus and/or Plasmid DNA were diluted to 298 ml in 0.01 M Tris pH 7.5, ImM EDTA. Forty ml 2M CaCl₂ was added followed by an equal volume of 2X HEPES buffered saline (lOg N-2-hydroxyethyl piperazine N' -2-ethanesulfonic acid (HEPES) , 16g NaCI, 0.74g KC1, 0.25g Na^PO^H , 2g dextrose per liter H₂0 and buffered with NaOH to pH 7.4) . The mixture was then incubated on ice for 10 minutes, and then added dropwise to an 80% confluent monolayer of CRFK cells growing in a 60 mm petri dish under 5 ml of medium (DME plus 5% fetal bovine serum) . The cells were incubated 4 hours at 37°C in a humidified incubator containing 5% C0₂. Media on the plates were aspirated, and cells were treated with 20% glycerol in 1XPBS (1.15g Na₂HP0₄, 0.2g KH₂P0₄, 0.8g NaCI, 0.2g KCl per liter₂ H 0) for one minute. The cells were washed three times with 5 ml of 1XPBS and then fed with 5ml of medium (DME plus 5% fetal bovine serum) . The cells were incubated at 37°C as above for 3-7 days until cytopathic effect from the virus was 50-100%. Virus was harvested as described above for the preparation of virus stocks. This stock was referred to as a transfection stock and was subsequently screened for recombinant virus by the

SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES.

HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS:

This method relies upon the homologous recombination between herpesvirus DNA and plasmid homology vector DNA which occurs in tissue culture cells co-transfected with these elements. From 0.1-1.0 mg of plasmid DNA containing foreign DNA flanked by appropriate herpesvirus cloned sequences (the homology vector) were mixed with approximately 0.3mg of intact herpesvirus DNA. The DNAs were diluted to 298 ml in 0.01 M Tris pH 7.5, ImM EDTA and transfected into CRFK cells according to the DNA TRANSFECTION FOR GENERATING RECOMBINANT VIRUS (see above) .

DIRECT LIGATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS:

Rather than using homology vectors and relying upon homologous recombination to generate recombinant virus, we have also developed the technique of direct ligation to engineer herpesviruses. In this instance, a cloned foreign gene did not require flanking herpesvirus DNA sequences but only required that it have restriction endonuclease sites available to cut out the foreign gene fragment from the plasmid vector. A compatible restriction enzyme was used to cut herpesvirus DNA. A requirement of the technique is that the restriction enzyme used to cut the herpesvirus DNA must cut at a limited number of sites. For FHV the restriction enzymes Sfil in S-FHV-010 is appropriate. Restriction endonuclease sites previously introduced into herpesviruses by other methods may also be used. The herpesvirus DNA is mixed with a 30-fold molar excess of plasmid DNA (typically 5mg of virus DNA to lOmg of plasmid DNA) , and the mixture is cut with the appropriate restriction enzyme. The DNA mixture is phenol extracted and ethanol precipitated to remove restriction endonucleases, and ligated together according to the ligation procedure detailed above. The ligated DNA mixture is then resuspended in 298 ml 0.01 M Tris pH 7.5, ImM EDTA and transfected into CRFK cells according to the DNA TRANSFECTION FOR GENERATING RECOMBINANT VIRUS (see above) .

PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS FROM SUBGENOMIC DNA FRAGMENTS: The ability to generate herpesviruses by cotransfection of cloned overlapping subgenomic fragments is known to those skilled in the art [26, 27] . If deletions and/or insertions are engineered directly into the subgenomic fragments prior to the cotransfection, this procedure results in a high frequency of viruses containing the genomic alteration, greatly reducing the amount of screening required to purify the recombinant virus. In the first step of this procedure deletions are introduced into separate viruses via homologous recombination with enzymatic marker genes as described below. The homology vector used in this step is constructed such that the enzymatic marker gene is flanked by a restriction endonuclease site that does not cut FHV in the region of the DNA to be deleted. In the second step a library of overlapping subgenomic fragments, capable of regenerating wild-type virus, is constructed from randomly sheared S-FHV-001 DNA. In the third step subgenomic fragments are cloned from each of the individual recombinant viruses containing attenuating deletion/marker gene insertions, which were generated in the first step. In each case the subcloned fragment corresponds in size and location to one of the wild-type subgenomic fragments constructed in the second step. This is accomplished by screening a library of randomly sheared recombinant virus DNA subclones with probes generated from the ends of the appropriate wild-type subgenomic fragment. The restriction endonuclease sites which had been engineered to flank the marker genes in the first step are now utilized to replace the marker genes in each subgenomic fragment with various foreign genes (such as FeLV env, FIV env, FIV gag, D. immi tis DiPLA2) . In the fourth step cotransfection of the appropriate overlapping wild type and deletion/insertion derived subgenomic fragments permits the generation of recombinant FHV viruses incorporating any desired combination of deletions and/or insertions.

SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING AN ENZYMATIC MARKER GENE:

When the E.coli β-galactosidase (lacZ) marker gene was incorporated into a recombinant virus the plaques containing recombinants were visualized by a simple assay. The enzymatic substrate was incorporated (300 mg/ml) into the agarose overlay during the plaque assay. For the lacZ marker gene the substrate BLUOGAL" (halogenated indolyl-?-D-galactoside, Life Technologies, Inc., Bethesda, MD) was used. Virus that expressed active marker enzyme turned blue. The blue plaques were then picked onto fresh CRFK cells, and the recombinant virus was purified by further blue plaque isolation. For recombinant virus construction in which the strategy requires that the enzymatic marker gene be removed, a subsequent assay involved plaque purifying white plaques from a background of parental blue plaques. In both cases viruses were typically purified with three to five sequential rounds of plaque purification.

CONSTRUCTION OF DELETION VIRUSES:

The strategy used to construct viruses containing genomic deletions involved the use of either homologous recombination and/or direct ligation techniques. Initially a virus was constructed via homologous recombination, in which the DNA to be deleted was replaced with a marker gene such as E. coli β- galactosidase (lacZ) . A second virus was then constructed in which the marker gene was subsequently deleted either by homologous recombination or via direct ligation. The advantage of this strategy is that both viruses may be purified by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The first virus is purified by picking blue plaques from a white plaque background, the second virus is purified by picking white plaques from a blue plaque background.

SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV USING BLACK PLAQUE ASSAYS: To analyze expression of foreign antigens expressed by recombinant FHV, onolayers of CRFK cells are infected with recombinant FHV, overlaid with nutrient agarose media and incubated for 1-2 days at 37°C. Once plaques have developed, the agarose overlay was removed from the dish, the monolayer rinsed" once with PBS, fixed with 100% methanol for 10 minutes at room temperature and the cells air dried. After re-hydrating the plate with PBS, the primary antibody was diluted to the appropriate dilution with PBS plus Blotto and incubated with the cell monolayer for 2 hours to overnight at room temperature. Unbound antibody was removed from the cells by washing four times with PBS at room temperature. The appropriate secondary antibody conjugate was diluted 1:500 with PBS and incubated with the cells for 2 hours at room temperature. Unbound secondary antibody was removed by washing the cells three times with PBS at room temperature. The monolayer was rinsed in color development buffer (lOOmM Tris pH 9.5/ lOOmM NaCI/ 5mM MgCl2), and incubated 10 minutes to overnight at room temperature with freshly prepared substrate solution (0.3 mg/ml Nitro Blue tetrazolium + 0.15 mg/ml 5-Bromo-4-Chloro-3-Indolyl Phosphatase in color development buffer) . the reaction was stopped by replacing the substrate solution with TE

(lOmM Tris, pH7.5/ 1 mM EDTA) . Plaques expressing the correct antigen stain black. ESTERN BLOTTING PROCEDURE: Samples of lysates and protein standards were run on a polyacrylamide gel according to the procedure of Laemnli [39] . After gel electrophoresis the proteins were transferred and processed according to Sambrook et al . [23] . The primary antibody was diluted 1:100 with 5% non-fat dry milk in Tris-sodium chloride, and Sodium Azide (TSA: 6.61g Tris-HCI, 0.97g Tris-base, 9. Og NaCI and 2. Og Sodium Azide per liter H₂0) . The secondary antibody was a alkaline phosphatase conjugate diluted 1:1000 with TSA.

ELISA ASSAY: Indirect Elisa assay was performed using standard techniques as described [35] .

SERUM-VIRUS NEUTRALIZATION ASSAY:

Sera were tested for FHV- and FeLV-specific antibodies by a microtiter technique. Test serum was inactivated for 30 minutes at 56°C. Duplicate 2-fold dilutions of test sera were made in 96-well microtitration plates with a 25 ml- pipettor. Equal volumes of virus suspension containing approximately 300 PFU were added to individual wells and the serum/virus mixtures were incubated at 37°C for 1 hour. 0.05 ml of a CRFK cell suspension containing approximately 4 X IO⁵ cells/ml was added to each well. The presence of antibody was indicated by the formation of a complete monolayer in 48 hours.

HOMOLOGY VECTOR 416-80.21B:

The homology vector 416-80.21B was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It contains a unique Hindlll site into which foreign genes were inserted. Upstream of the unique Hiπdlll site is an approximately 1417 base pair fragment of FHV DNA which includes all of the gl gene coding sequence. Downstream of the unique Hindlll site is an approximately 5200 base pair fragment of FHV DNA which includes part of the terminal repeat sequence. A detailed description of the plasmid is given in Figures 4A-4B. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65 (Promega Corp., Madison, WI) . Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment

HOMOLOGY VECTOR 644-09.A1:

The homology vector 644-09.A1 was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It contains a unique Hindlll site into which foreign genes were inserted. A detailed description of the plasmid is given in Figures lOA-lOC. It was constructed from the indicated DNA sources utlilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2958 base pair Asp7l8I restriction endonuclease fragment of a pSP18/pSP19 fusion such that the multiple cloning site is EcoRI / Sacl /Asp! 181 /Sacl /EcoRI . Fragment 1 is an approximately 1415 base pair Asp718I to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 2205 base pair Sail to Asp718I subfragment of FHV EcoRI E fragment .

HOMOLOGY VECTOR 644-09.B2:

The homology vector 644-09.B2 was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It contains a unique

site into which foreign genes were inserted. A detailed description of the plasmid is given in Figures 11A-11B. It was constructed from the indicated DNA sources utlilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2958 base pair Asp718I restriction endonuclease fragment of a pSP18/pSP19 fusion such that the multiple cloning site is EcoRI/SacI/Asp718I/SacI/EcoRI. Fragment 1 is an approximately 1415 base pair Asp718I to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 2220 base pair Sail to Asp718I subfragment of FHV EcoRI E fragment .

HOMOLOGY VECTOR 478-17.IT:

The homology vector 478-17.IT was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli 3-galactosidase gene and a pseudorabies virus glycoprotein C-feline leukemia virus glycoprotein 70 (PRV gC-FeLV g70) envelope fusion polypeptide flanked by FHV DNA. The FeLV g70 envelope epitope (FEE #1) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-MGPNLVLPDQKPPS-COOH; Ref. 21; Figure 2, SEQ ID NO: 8) . The homology vector 478- 17. IT contains eight copies of FeLV g70 envelope epitope #1 fused in frame internal to the PRV gC polypeptide. The foreign DNA was inserted into a unique Sail site in the FHV homology vector 416-80.21. Upstream of the unique Sail site is an approximately 1417 base pair fragment of FHV DNA that includes all of the FHV gl gene coding sequence. Downstream of the unique Sail site is an approximately 5200 base pair fragment of FHV DNA that includes the unique short sequence and the terminal repeat sequence. A detailed description of the plasmid 478-17.IT is given in Figures 7A-7E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 943 base pair iVcol to Notl subfragment of PRV BamHI #2 [29] . Fragment 4 is an approximately 408 base pair fragment of FeLV g70 envelope gene DNA (8 copies of FeLV g70 envelope epitope #1; Figure 2) . Fragment -5 is an approximately 1116 base pair X ol to Ncol subfragment of PRV BamHI #2 and #9 [29] . Fragment 6 is an approximately 543 base pair iVdel to Kpnl subfragment of PRV BaπiHI #7 [29] . Fragment 7 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10 [29] . Fragment 8 is an approximately 3388 base pair BamHI to Ball subfragment of pJF751 [28] . Fragment 9 is an approximately 784 base pair Smal to Smal subfragment of HSV-1 BamHI Q [30] . Fragment 10 is an approximately 221 base pair Kpnl to Sail subfragment of PRV BamHI #7 [29] . Fragment 11 is an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment

HOMOLOGY VECTOR 478-10.11:

The homology vector 478-10.11 was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli _/S-galactosidase gene and a PRV gC-FeLV g70 envelope fusion polypeptide flanked by FHV DNA. The FeLV g70 envelope epitope (FEE #1) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-MGPNLVLPDQKPPS-COOH; Ref. 21; Figure 2, SEQ ID NO: 8)^".^' The homology vector 478-10.11 contains four copies of FeLV g70 envelope epitope #1 fused in frame internal to the PRV gC polypeptide. The foreign genes were inserted into a unique Sail site in the FHV homology vector 416-80.21. A detailed description of the plasmid 478-10.11 is given in Figures 7A-7E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment (of the FHV Sail B fragment. Fragment 2 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 943 base pair Wcol to Notl subfragment of PRV BamHI #2 [29] . Fragment 4 is an approximately 204 base pair fragment of FeLV g70 envelope gene DNA (4 copies of FeLV g70 envelope epitope #1; Figure 2) . Fragment 5 is an approximately 1116 base pair Xhol to Wcol subfragment of PRV BamHI #2 and #9 [29] . Fragment 6 is an approximately 543 base pair Ndel to Kpnl subfragment of

PRV BamHI #7 [29] . Fragment 7 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10

[29] . Fragment 8 is an approximately ^• 3388 base pair BamHI to Ball subfragment of pJF751 [28] . Fragment 9 is an approximately 784 base pair Smal to Smal subfragment of HSV-1 BamHI Q [30] . Fragment 10 is an approximately 221 base pair Kpnl to Sail subfragment of PRV BamHI #7 [29] . Fragment 11 is an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment

HOMOLOGY VECTOR 411-91.01A:

The homology vector 411-91.01A was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli jβ-galactosidase gene and a PRV gC-FeLV g70 envelope fusion polypeptide flanked by FHV DNA. The FeLV g70 envelope epitope (FEE #1) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-MGPNLVLPDQKPPS-COOH; Ref. 21; Figure 2, SEQ ID NO: 8) . The homology vector 411-91.01A contains four copies of FeLV g70 envelope epitope #1 fused in frame internal to the PRV gC polypeptide. The foreign genes were inserted into Pstl and Sail sites in the FHV homology vector 416-80.21. A detailed description of the plasmid 411-91.01A is given in Figures 3A-3D. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 3006 base pair BamHI to PvuII subfragment of pJF751 [28] . Fragment 4 is an approximately 204 base pair of FeLV g70 envelope gene DNA (4 copies of FeLV g70 envelope epitope #1; Figure 2) . Fragment 5 is an approximately 751 base pair Wdel to Sail subfragment of PRV BamHI #7 [29] . Fragment 6 is an approximately^• 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment.

HOMOLOGY VECTOR 411-91.01B:

The homology vector 411-91.01B was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli lacZ-FeLV g70 envelope fusion polypeptide flanked by FHV DNA. The FeLV g70 envelope epitope (FEE #1) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-MGPNLVLPDQKPPS-COOH) (Ref. 21; Figure 2, SEQ ID NO: 8) , and the FeLV g70 envelope epitope (FEE #3) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-AKLRERLKEREQLF-COOH) (Ref. 21; Figure 2, SEQ ID NO: 10) . The homology vector 411-91.01A contains four copies of FEE #1, one copy of FEE #3, one copy of FEE #1 and one copy of FEE #3 fused in frame to the carboxy terminus of the E. coli β- galactosidase (lacZ) polypeptide. The foreign gene was inserted into Pstl and Sail sites in the FHV homology vector 416-80.21. A detailed description of the plasmid 411-91.01B is given in Figures 3A-3D. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 3006 base pair BamHI to PvuII subfragment of pJF751 [28] . Fragment 4 is an approximately 357 base pair assembly of the FeLV g70 envelope gene (a concatemer of four copies of FEE #1, one copy of FEE #3, one copy of FEE #1, and one copy of FEE #3; Figure 2) . Fragment 5 is an approximately 751 base pair Wdel to Sail subfragment of PRV BamHI #7 [29] . Fragment 6 is an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment.

HOMOLOGY VECTOR 510-36.1C: The homology vector 510-36. IC was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli _/S-galactosidase gene and a hepatitis B core antigen-FeLV g70 envelope fusion polypeptide flanked by FHV DNA. The FeLV g70 envelope epitope (FEE #1) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-MGPNLVLPDQKPPS-COOH; Ref. 21; Figure 2, SEQ ID NO: 8) . The homology vector 510- 36.1C contains four copies of FeLV g70 envelope epitope #1 fused in frame at the carboxy terminus of the hepatitis B core antigen polypeptide. The foreign genes were inserted into Pstl and Sail sites in the FHV homology vector 416-80.21. A detailed description of the plasmid 510-36.IC is given in Figures 8A-8E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 204 base pair fragment of FeLV g70 envelope gene DNA (4 copies of FEE #1; Figure 2) . Fragment 4 is an approximately 590 base pair BamHI to Kpnl fragment of hepatitis B core antigen DNA. Fragment 5 is an approximately 543 base pair Wdel to Kpnl subfragment of PRV BamHI #7

[29] . Fragment 6 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10 [29] . Fragment 7 is an approximately 3388 base pair ^■ BamHI to Ball subfragment of pJF751 [28] . Fragment 8 is an approximately 784 base pair Smal to Smal subfragment of

HSV-1 BamHI Q [30] . Fragment 9 is an approximately 221 base pair Kpnl to Sail subfragment of PRV BamHI #7

[29] . Fragment 10 is an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment.

HOMOLOGY VECTOR 510-36.2F:

The homology vector 510-36.2F was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli β-galactosidase gene and a hepatitis B core antigen-FeLV g70 envelope fusion polypeptide flanked by FHV DNA. The FeLV g70 envelope epitope (FEE #1) polypeptide consists of a 14 amino acid antigenic epitope (NH₂-MGPNLVLPDQKPPS-COOH; Ref. 21; Figure 2, SEQ ID NO: 8) . The homology vector 510- 36.2F contains eight copies of FeLV g70 envelope epitope #1 fused in frame at the carboxy terminus of the hepatitis B core antigen polypeptide. The foreign genes were inserted into Pstl and Sail sites in the FHV homology vector 416-80.21. A detailed description of the plasmid 510-36.2F is given in Figures 8A-8E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 408 base pair fragment of FeLV g70 envelope gene DNA (8 copies of FEE #1; Figure 2) . Fragment 4 is an approximately 590 base pair BamHI to Kpnl fragment of hepatitis B core antigen gene DNA. Fragment 5 is an approximately 543 base pair iVdel to Kpnl subfragment- of PRV BamHI #7 [29] . Fragment 6 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10 [29] . Fragment 7 is an approximately 3388 base pair BamHI to Ball subfragment of pJF751 [28] . Fragment 8 is an approximately 784 base pair Smal to Smal subfragment of HSV-1 BamHI Q [30] . Fragment 9 is an approximately 221 base pair Kpnl to Sail subfragment of PRV BamHI #7

[29] . Fragment 10 is an approximately 5200 base pair

Sail to PvuII subfragment of the FHV EcoRI E fragment

HOMOLOGY VECTOR 416-88.2L:

The homology vector 416-88.2L was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli β-galactosidase gene flanked by FHV DNA. The foreign gene was inserted into a Pstl site in the FHV homology vector 416-80.21. A detailed description of the plasmid 416-88.2L is given in Figures 5A-5C. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2817 base pair Sacl to PvuII restriction endonuclease fragment of a pSP65. Fragment 1 is an approximately 1417 base pair Sacl to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10 [29] . Fragment 3 is an approximately 3388 base pair BamHI to Ball subfragment of pJF751 [28] . Fragment 4 is an approximately 751 base pair Ndel to Sail subfragment of PRV BamHI #7 [29] . Fragment 5 is an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment.

HOMOLOGY VECTOR 639-85.IH:

The homology vector 639-85. IH was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli /3-galactosidase gene and a Dirofilaria immi tis DNA clone encoding a 22 kd immunogenic polypeptide (DiPLA2) flanked by FHV DNA. The foreign genes were inserted into Notl site in the FHV homology vector 644-09.B2. A detailed description of the plasmid 639-85.IH is given in Figures 9A-9E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2958 base pair Asp718I to Asp718I restriction endonuclease fragment of a pSP18/19. Fragment 1 is an approximately 1415 base pair Asp718I to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 179 base pair Pstl to EcoRV subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 3 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10 [29] . Fragment 4 is an approximately 3006 base pair BamHI to PvuII subfragment of pJF751 [28] . Fragment 5 is an approximately 751 base pair Wdel to Sail subfragment of PRV BamHI #7 [29] . Fragment 6 is an approximately 977 base pair EcoRV to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 7 is an approximately 618 base pair EcoRI to Xhol fragment of the D. immi tis DiPLA2 gene [34] . Fragment 8 is an approximately 2205 base pair Sail to Asp718I subfragment of the FHV EcoRI E fragment .

HOMOLOGY VECTOR 669-42.04:

The homology vector 669-42.04 was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli -galactosidase gene and a feline immunodeficiency virus (FIV) protease

(gag) gene flanked by FHV DNA. A portion of the FIV gag gene was synthesized by the polymerase chain reaction

(PCR) . The foreign genes were inserted into the iVotl site in the FHV homology vector 644-09.B2. A detailed description of the plasmid 669-42.04 is given in Figures 12A-12E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2958 base pair Asp718I to Asp718I restriction endonuclease fragment of a pSP18/19. Fragment 1 is an approximately 1415 base pair Asp718I to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 221 base pair Sail to Kpnl subfragment of PRV BamHI #7 [29] . Fragment 3 is an approximately 1154 base pair Pstl to Avail subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 4 is an approximately 1853 base pair Bgrlll to iVcol fragment of the FIV gag gene [32] . The 3' end of the gene is an approximately 1360 base pair Xmal to Ncol subfragment of the FIV PPR genomic cDNA. The 5' end of the gene (an approximately 560 base pair Bglll to Xmal fragment) was synthesized by PCR. The template for the PCR reaction was FIV strain PPR genomic cDNA [32] . The upstream primer 03 (5' -GCGAGATCTGAAAATGGCCATTAAGAGATG-3' ; SEQ ID NO: 3) was synthesized to correspond to DNA from the 5' end of the FIV gag gene starting at nucleotide 653 of FIV strain PPR genomic cDNA (amino acid 10 of FIV gag protein) and to introduce a Bglll site at the 5' end. The downstream primer 04 (5' -GTGCTGCAGTAAAATAGGG-3 ' ; SEQ ID NO: 4) was synthesized to correspond to DNA starting at nucleotide 1323 of FIV PPR genomic cDNA and is positioned downstream of an Xmal site at nucleotide 1207. Fragment 5 is an approximately 784 base pair Smal to Smal subfragment of the HSV-1 BamHI Q fragment [30] . Fragment 6 is an approximately 543 base pair Kpnl to Wdel subfragment of PRV BamHI #7 [29] . Fragment 7 is an approximately 3006 base pair PvuII to BamHI subfragment of pJF751 [28] . Fragment 8 is an approximately 423 base pair BamHI to Sail subfragment of PRV BamHI #10 [29] . Fragment 9 is an approximately 2205 base pair Sail to Asp718I subfragment of the FHV EcoRI E fragment.

HOMOLOGY VECTOR 725-26.A10:

The homology vector 725-26.A10 was constructed for the purpose of deleting a portion of the gE coding region from the feline herpesvirus and inserting a foreign DNA. It incorporates an E. coli β-galactosidase gene and a feline immunodeficiency virus (FIV) envelope (env) gene flanked by FHV DNA. The FIV env gene was synthesized by the polymerase chain reaction (PCR) . The foreign genes were inserted into a Wbtl site in the FHV homology vector 644-09.B2 (Figures 13A-13E) . A detailed description of the plasmid 639-85.IH is given in Figures 9A-9E. It was constructed from the indicated DNA sources utilizing standard recombinant DNA techniques [22, 23] . The plasmid vector is derived from an approximately 2958 base pair Asp718I to Asp718I restriction endonuclease fragment of a pSP18/l9. Fragment 1 is an approximately 1415 base pair Asp718I to Smal subfragment of the FHV Sail B fragment. Fragment 2 is an approximately 423 base pair Sail to BamHI subfragment of PRV BamHI #10 [29] . Fragment 3 is an approximately 3006 base pair BamHI to PvuII subfragment of pJF751 [28] . Fragment 4 is an approximately 751 base pair Ndel to Sail subfragment of- PRV BamHI #7 [29] . Fragment 5 is an approximately 784 base pair Smal to Smal subfragment of the HSV-1 BamHI Q [30] . Fragment 6 is an approximately 2564 base pair BamHI to BamHI fragment of the FIV env gene [32] synthesized by PCR. The template for the PCR reaction was FIV strain PPR genomic cDNA [32] . The upstream primer 10/93.21BW (5' -GCCCGGATCCTATGGCAGAAGGGTTTGCAGC- 3' ; SEQ ID NO: 5) was synthesized corresponding to the 5' end of the FIV env gene starting at nucleotide 6263 of FIV strain PPR genomic cDNA, and the procedure introduced a BamHI site at the 5' end. The BamHI site was destroyed during the cloning of the PCR fragment . The downstream primer 10/93.20BW (5'- CCGTGGATCCGGCACTCCATCATTCCTCCTC-3' ; SEQ ID NO: 6) was synthesized corresponding to the 3' end of the FIV env gene starting at nucleotide 8827 of FIV PPR genomic cDNA, and the procedure introduced a BamHI site at the 3' end. Fragment 7 is an approximately 1154 base pair Avail to Pstl subfragment of the HCMV 2.1 kb Pstl E fragment [31] . Fragment 8 is an approximately 2205 base pair Sail to Asp718I subfragment of the FHV EcoRI E fragment .

Example 1: Recombinant feline herpesvirus (FHV) containing a deletion of the entire gE gene and an insertion of a foreign DNA sequence into that site will replicate in cats and is useful as a vaccine. The ability to isolate a gE-deleted FHV confirms that the FHV gE gene

(SEQ ID NOs: 1 and 2) is non essential for replication of the recombinant FHV. Recombinant FHV expressing foreign genes for viral, bacterial or parasite antigens protect against disease in dogs and cats. Recombinant FHV which was isolated contains a deletion of the gE gene within the unique short and an insertion of a foreign gene into that gE site or any nonessential site will replicate in cats and is useful as a vaccine.

Recombinant FHV containing a deletion of the gE gene within the unique short and an insertion of a foreign gene into that site is useful as a vaccine. Recombinant FHV expressing foreign genes for viral, bacterial, or parasite antigens is useful as a vaccine to protect against disease in cats, dogs, humans, horses, cattle, swine and poultry.

Homology vectors 416-80.21, 644-09.Al and 644-09.B2 are deleted for the entire gE gene and are useful in the present invention for insertion and expression of foreign genes in recombinant FHV. Homology vectors 416- 80.21, 644-09.Al and 644-09.B2 carry a 1638 base pair deletion of the gE gene from the Smal site in the FHV Sail B fragment to the Sail site in the FHV EcoRI E fragment (see Figures 4, 10 and 11) . The sequence of the gE gene (SEQ ID NOs: 1 and 2) includes the coding sequence for the 531 amino acid open reading frame of the gE gene and also includes the Smal site in the FHV Sail B fragment and the Sail site in the FHV EcoRI E fragment which define the endpoints of the deletion of the gE gene. To one side of the FHV gE deletion, homology vectors 416-80.21, 644-09.Al and 644-09.B2 contain an approximately 1415 base pair Asp718I to Smal subfragment of FHV Sail B containing the entire coding sequence of the gl gene (370 amino acids) . To the opposite side of the FHV gE deletion, homology vector 416-80.21 contains an approximately 5200 base pair Sail to PvuII subfragment of the FHV EcoRI E fragment which contains unique short and terminal repeat sequences. To the opposite side of the FHV gE deletion in homology vectors 644-09.Al and 644-09.B2, an approximately 2205 base pair Sail to Asp718I subfragment of the FHV EcoRI E fragment contains unique short and terminal repeat sequences.

Example 2 :

Recombinant FHV containing a deletion of the gG gene within the unique short and an insertion of a foreign gene into that site will replicate in cats and is useful as vaccine. Recombinant FHV expressing foreign genes for viral, bacterial, or parasite antigens is useful as a vaccine to protect against disease in cats, dogs, humans, horses, cattle, swine and poultry.

The ability to isolate a gG-deleted FHV confirms that the FHV gG gene is non-essential for replication of the recombinant FHV. Utilizing the sequence of the gG (SEQ ID NOs : 71 and 72), a homology vector containing a deletion of the gG gene and an insertion of a foreign gene is constructed. A recombinant FHV is isolated utilizing the gG deleted homology vector and virus S- FHV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS.

Example 3 :

Recombinant FHV containing a deletion of the gl gene within the unique short or a deletion in the repeat region, and an insertion of a foreign gene into that site will replicate in cats and is useful as a vaccine. Recombinant FHV expressing foreign genes for viral, bacterial, or parasite antigens is useful as a vaccine to protect against disease in cats, dogs, humans, horses, cattle, swine and poultry.

The ability to isolate a gl-deleted FHV or repeat- deleted FHV confirms that the FHV gl gene or FHV repeat sequence is non-essential for replication of the recombinant FHV. Utilizing the sequence of the gl gene or repeat sequence, a homology vector containing a deletion of the gl gene or repeat sequence, and an insertion of a foreign gene is constructed. A recombinant FHV is isolated utilizing the gl deleted homology vector or the repeat-deleted homology vector, and virus S-FHV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS.

Recombinant FHV containing a deletion of the gE gene and a deletion of the gG and/or gl genes within the unique short and an insertion of a foreign gene into a nonessential site and is useful as a vaccine to protect against disease in cats, dogs, humans, horses, cattle, swine and poultry.

Example 4:

Recombinant feline herpesvirus express antigenic epitopes of the feline leukemia virus envelope gene

(gag, pol, env) fused to the E. coli lacZ gene in animals. Recombinant viruses S-FHV-003, S-FHV-004 and S-FHV-006 are each useful as a vaccine in cats against feline leukemia and feline rhinotracheitis.

Example 4A S-FHV-003 S-FHV-003 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. A fusion gene containing four copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2, SEQ ID NOs: 7 and 8) is fused in the correct translational reading frame to the 3' end of the E. coli lacZ gene (_/S-galactosidase) . The FeLV env-lacZ fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-003 was derived from S-FHV-000 (ATCC No. 636) . This was accomplished utilizing the homology vector 411-91.01A (see Materials and Methods) and virus S-FHV- 000 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-003. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-lacZ fusion gene and the deletion of the 1638 base pair gE gene.

S-FHV-003 was assayed for expression of FeLV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera was shown to react specifically with S-FHV-003 plaques and not with S-FHV- 000 negative control plaques. Every viral plaque of S- FHV-003 that was analyzed from all stages of propagation reacted with the antiserum indicating that the virus was stably expressing the FeLV foreign gene. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines .

To confirm the expression of the S-galactosidase-FeLV env fusion gene product, cells were infected with S- FHV-003 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal mouse anti FeLV EE#1 peptide antisera was used to detect expression of β- galactosidase-FeLV envelope fusion protein. The lysate from S-FHV-003 infected cells exhibited a band at approximately 150 kd which is the expected size of the 3-galactosidase-FeLV env fusion protein.

Example 4B S-FHV-004

S-FHV-004 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. A fusion gene containing four copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2, SEQ ID NOs: 7 and 8) , one copy of FeLV EE #3 (Figure 2, SEQ ID NOs: 9 and 10) , one copy of FeLV EE #1, and one copy of FeLV EE #3 are fused in the correct translational reading frame to the 3' end of the E. coli lacZ gene (β- galactosidase) . The FeLV env-lacZ fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-004 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 411-91.01B (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-004. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-lacZ fusion gene and the deletion of the 1638 base pair gE gene.

Table 1

Elisa Assay and Serum-Virus Neutralizing Titers from Cats Vaccinated with S-FHV-003 and S-FHV-004

Vaccine Elisa FeLV FHV crp70^» SN^a SNa Day 20 Day 29 Day 29 Day 69 Day 20 Day 29 Day 59

S-FHV- 0/4 4/4 1/4 0/4 0/4 0/4 0/4

003

S-FHV- 0/4 3/4 1/4 0/4 0/4 3/4 3/4 004 ^a Number of cats with a positive Elisa titer (0.05 or greater) or SN titer (1:4 or greater) per 4 cats total per group.

Table 1 shows the results of vaccinating cats with recombinant FHV S-FHV-003 or S-FHV-004 (4 cats in each group) . Cats were vaccinated intramuscularly with approximately 1 X IO⁶ PFU of S-FHV-003 or S-FHV-004 on Day 0, Day 20, and Day 59. Sera from cats bled on days 0, 29, 59 and 69 were assayed for antibodies to FeLV gp70 protein. ELISA ASSAY detected FeLV gp70 -specif ic antibodies in the serum of S-FHV-003 and S-FHV-004 vaccinated cats. The SERUM-VIRUS NEUTRALIZATION ASSAY

(SN) titers against FeLV were low in the serum of S- FHV-004 vaccinated cats, while SN titers against FHV were 1:8 or greater at day 29 and day 59. An SN titer greater than or -equal to 1:8 is necessary to resist challenge with FHV. The results indicate that S-FHV-003 and S-FHV-004 are useful to express a foreign gene in vivo, such as the FeLV gp70 antigenic epitope. The results also^" indicate that S-FHV-004 is useful as a vaccine against feline rhinotracheitis.

To confirm the expression of the _/8-galactosidase-FeLV env fusion gene product, cells were infected with S- FHV-004 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal mouse anti FeLV EE#1 peptide antisera was used to detect expression of β- galactosidase-FeLV envelope fusion protein. The lysate from S-FHV-004 infected cells exhibited a band at approximately 150 kd which is the expected size of the _/S-galactosidase-FeLV env fusion protein.

Example 4C S-FHV-006

S-FHV-006 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. A fusion gene containing four copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2 SEQ ID NOs: 7 and 8) is fused in the correct translational reading frame to the 3' end of the E. coli lacZ gene (_/β-galactosidase) . The FeLV env-lacZ fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-006 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 411-91.01A (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-006. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-lacZ fusion gene and the deletion of the 1638 base pair gE gene.

S-FHV-006 was assayed for expression of FeLV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera was shown to react specifically with S-FHV-006 plaques and not with S-FHV- 000 negative control plaques. Every viral plaque of S- FHV-006 that was analyzed from all stages of propagation reacted with the antiserum indicating that the virus was stably expressing the FeLV foreign gene. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

To confirm the expression of the _/S-galactosidase-FeLV env fusion gene product, cells were infected with S-

FHV-006 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis.

The gel was blotted and analyzed using the WESTERN

BLOTTING PROCEDURE. Polyclonal mouse- anti FeLV EE#1 peptide antisera was used to detect expression of β- galactosidase-FeLV envelope fusion protein. The lysate from S-FHV-006 infected cells exhibited a band at approximately 150 kd which is the expected size of the 3-galactosidase-FeLV envelope fusion protein.

Example 5:

Recombinant feline herpesviruses express antigenic epitopes of the feline leukemia virus envelope gene fused to the pseudorabies virus glycoprotein C (PRV gC) gene in animals. Recombinant viruses S-FHV-007, S-FHV- 009 and S-FHV-012 are each useful as a vaccine in cats against feline leukemia and feline rhinotracheitis.

Example 5A S-FHV-007

S-FHV-007 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. A fusion gene containing six copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2 SEQ ID NOs: 7 and 8) is fused in the correct translational reading frame within the PRV gC gene. The six copies of FeLV EE#1 are in frame between amino acids 1 to 313 and amino acids 421 to 467 of the PRV glycoprotein C. The FeLV env-PRV gC fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-007 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 478-17.IT (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-007. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-PRV gC fusion gene and the deletion of the 1638 base pair gE gene. The homology vector 478-17. IT used to construct S-FHV- 007 contains eight copies of FeLV EE#1. However in the course of homologous recombination and purification of S-FHV-007, the virus contains six copies of FeLV EE#1 fused to the PRV gC gene. This was confirmed by DNA SEQUENCE ANALYSIS of the S-FHV-007 virus. S-FHV-007 was assayed for expression of FeLV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera was shown to react specifically with S-FHV-007 plaques and not with S-FHV- 000 negative control plaques. Every viral plaque of S- FHV-007 that was analyzed from all stages of propagation reacted with the antisera indicating that the virus was stably expressing the FeLV envelope peptide. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

To confirm the expression of the FeLV env-PRV gC fusion gene product, cells were infected with S-FHV-007 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal goat 282 antiserum reactive against denatured PRV gC antigen [33] were used to detect expression of FeLV env-PRV gC fusion protein. The lysate from S-FHV-007 infected cells exhibited a viral specific band at approximately 90 to 100 kd that is the expected size of the FeLV env-PRV gC fusion protein.

Example 5B S-FHV-009

S-FHV-009 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. A fusion gene containing eight copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2, SEQ ID NOs: 7 and 8) is fused in the correct translational reading frame within the PRV gC gene. The eight copies of FeLV EE#1 are in frame between amino acids 1 to 313 and amino acids 421 to 467 of the PRV glycoprotein C. The FeLV env-PRV gC fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-009 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 478-17.IT (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-009. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-PRV gC fusion gene and the deletion of the 1638 base pair gE gene.

S-FHV-009 was assayed for expression of FeLV-and PRV- specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal goat 282 antiserum reactive against denatured PRV gC antigen [33] were shown to react specifically with S- FHV-009 plaques and not with S-FHV-000 negative control plaques. Every viral plaque of S-FHV-009 that was analyzed from all stages of propagation reacted with the antisera indicating that the virus was stably expressing the FeLV envelope peptide and the PRV glycoprotein C. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

To confirm the expression of the FeLV env-PRV gC fusion gene product, cells were infected with S-FHV-009 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal goat 282 antiserum reactive against denatured PRV gC antigen [33] were used to detect expression of FeLV env-PRV gC fusion protein. The lysate from S-FHV-009 infected cells exhibited a band at approximately 90 to 100 kd which is the expected size of the FeLV env-PRV gC fusion protein.

Example 5C S-FHV-012

S-FHV-012 is a recombinant feline herpesvirus that has a deletion of the gE gene and the insertion of a foreign gene at the gE deletion site. A fusion gene containing four copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2, SEQ ID NOs: 7 and 8) is fused in the correct translational reading frame within the PRV gC gene. The four copies of FeLV EE#1 are in frame between amino acids 1 to 313 and amino acids 421 to 467 of the PRV glycoprotein C. The FeLV env-PRV gC fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-012 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 478-10.11 (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-012. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-PRV gC fusion gene and the deletion of the 1638 base pair gE gene.

S-FHV-012 was assayed for expression of FeLV-and PRV- specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal goat 282 antiserum reactive against denatured PRV gC antigen [33] were shown to react specifically with S- FHV-012 plagues artd not with S-FHV-000 negative control plaques. Every viral plaque of S-FHV-012 that was analyzed from all stages of propagation reacted with the antisera indicating that the virus was stably expressing the FeLV envelope peptide and the PRV glycoprotein C. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

Example 6:

Recombinant feline herpesviruses express antigenic epitopes of the feline leukemia virus envelope gene fused to the hepatitis B virus core antigen gene in animals. Recombinant viruses S-FHV-008 and S-FHV-011 are each useful as a vaccine in cats against feline leukemia and feline rhinotracheitis.

Example 6A S-FHV-008

S-FHV-008 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. A fusion gene containing four copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2, SEQ ID NOs: 7 and 8) is fused with the correct translational reading frame to the 5' end of the hepatitis B core antigen gene. The FeLV env-hepatitis B core antigen fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-008 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 510-36.1C (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-008. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-hepatitis B core antigen fusion gene and the deletion of the 1638 base pair gE gene.

S-FHV-008 was assayed for expression of FeLV- and hepatitis B-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal rabbit anti-hepatitis B core antigen antisera were shown to react specifically with S-FHV- 008 plaques and not with S-FHV-000 negative control plaques. Every viral plaque of S-FHV-008 that was analyzed from all stages of propagation reacted with the antiserum indicating that the virus was stably expressing the FeLV env-hepatitis B core antigen fusion foreign gene. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

To confirm the expression of the FeLV env-hepatitis B core antigen fusion gene product, cells were infected with S-FHV-008 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal rabbit anti hepatitis B core antigen antisera were used to detect expression of FeLV env-hepatitis B core antigen fusion protein. The lysate from S-FHV-008 infected cells exhibited a band at approximately 32 kd which is the expected size of the FeLV env (4 copies of FeLV EE#D- hepatitis B core antigen fusion protein.

Example 6B S-FHV-011

S-FHV-011 is a recombinant feline herpesvirus that has a deletion of the gE gene within the unique short region and the insertion of two foreign genes at the gE deletion site. A fusion gene containing eight copies of the feline leukemia virus envelope epitope #1 (FeLV EE #1; Figure 2, SEQ ID NOs: 7 and 8) is fused with the correct translational reading frame to the 5' end of the hepatitis B core antigen gene. The FeLV env- hepatitis B core antigen fusion gene is under the transcriptional control of the HCMV immediate early promoter.

S-FHV-011 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 510-36.2F (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-011. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FeLV env-hepatitis B core antigen fusion gene and the deletion of the 1638 base pair gE gene.

S-FHV-011 was assayed for expression of FeLV- and hepatitis B-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal rabbit anti-hepatitis B core antigen antisera were shown to react specifically with S-FHV- 011 plaques and not with S-FHV-000 negative control plaques. Every viral plaque of S-FHV-011 that was analyzed from all stages of propagation reacted with the antiserum indicating that the virus was stably expressing the FeLV env-hepatitis B core antigen fusion foreign gene. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

To confirm the expression of the FeLV env-hepatitis B core antigen fusion gene product, cells were infected with S-FHV-011 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal mouse anti FeLV EE#1 peptide antisera and polyclonal rabbit anti hepatitis B core antigen antisera were used to detect expression of FeLV env-hepatitis B core antigen fusion protein. The lysate from S-FHV-011 infected cells exhibited a band at approximately 40 kd which is the expected size of the FeLV env (8 copies of FeLV EE#1) -hepatitis B core antigen fusion protein.

Example 7; Recombinant feline herpesvirus express the E. coli lacZ (/3-galactosidase) gene in animals. Recombinant viruses S-FHV-005 and S-FHV-010 are each useful as a vaccine in cats against feline rhinotracheitis.

Example 7A S-FHV-005

S-FHV-005 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of a foreign gene at the gE deletion site. The foreign gene is an E. coli lacZ (_/S-galactosidase) gene under the transcriptional control of the PRV gX promoter.

S-FHV-005 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 416-88.2L (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-005. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure.- This analysis confirmed the insertion of the E. coli lacZ ( β- galactosidase) genes and the deletion of the 1638 base pair gE gene.

Example 7B S-FHV-010

S-FHV-010 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of an unique Sfil restriction endonuclease site at the gE deletion site.

S-FHV-010 was derived from S-FHV-005. This was accomplished utilizing the virus S-FHV-005 which was digested with Sfil and religated to remove the PRV gX promoter-E. coli lacZ marker cassette leaving a unique Sfil restriction endonuclease site within the FHV gE deletion site. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final-result of white plaque purification was the recombinant virus designated S-FHV-010. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the Sfil restriction endonuclease site and the deletion of the 1638 base pair gE gene.

Example 8 S-FHV-014

S-FHV-014 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of two foreign genes at the gE deletion site. The Dirofilaria immi tis 22 kD (DiPLA2) gene is under the transcriptional control of the HCMV immediate early promoter and the E. coli lacZ (_/β-galactosidase) gene is under the transcriptional control of the PRV gX promoter.

S-FHV-014 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 639-85.1H (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-014. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the D. immi tis 20/22 kD (DiPLA2) and the E. coli lacZ (/S-galactosidase) genes and the deletion of the 1638 base pair gE gene.

S-FHV-014 was assayed for expression of Dirofilaria immi tis 22kD (DiPLA2) -specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT FHV. Rabbit anti-recombinant 22kd (Dipla2) antisera was shown to react specifically with S-FHV-014 plaques and not with S-FHV-000 negative control plaques. Every viral plaque of S-FHV-014 that was analyzed from all stages of propagation reacted with the antiserum indicating that the virus was stably expressing the D. immi tis 22kD (DiPLA2) foreign gene. The assays described here were carried out in CRFK cells, indicating that CRFK cells would be a suitable substrate for the production of FHV recombinant vaccines.

To confirm the expression of the D. immi tis 20/22kD DiPLA2 gene product, cells were infected with S-FHV-014 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal dog convalescent anti -Dirofilaria immi tis antisera was used to detect expression of D. immi tis 20/22kD DiPLA2 protein. The lysate from S-FHV- 014 infected cells exhibited bands at approximately 26kd, 28kd, 32kd which correspond to glycosylated forms of the D. immi tis 20/22kD DiPLA2 protein. Rabbit anti- recombinant Dipla2 antisera reacted very strongly with a 17 kd protein on a Western blot. S-FHV-014 is useful as a vaccine in dogs and cats against heartwor disease caused by the D. immi tis parasite. Cats vaccinated with S-FHV-014 demonstrated significant protection from infection after challenge with the D. immi tis parasite compared to unvaccinated control cats.

Example 9: Recombinant feline herpesviruses express feline immunodeficiency virus (FIV) and the E. coli lacZ ( β- galactosidase) genes in animals. Recombinant viruses S- FHV-016 and S-FHV-017 are useful as a vaccine in cats against feline rhinotracheitis and disease caused by feline immunodeficiency virus.

Example 9A S-FHV-016

S-FHV-016 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of two foreign genes at the gE deletion site. The FIV gag (p50; protease) gene is under the transcriptional control of the HCMV immediate early promoter and the E. coli lacZ (_/β-galactosidase) gene is under the transcriptional control of the PRV gX promoter.

S-FHV-016 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 669-42.04 (see Materials and Methods) and virus S-FHV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-016. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FIV gag (p50) gene and the E. coli lacZ (3-galactosidase) gene and the deletion of the 1638 base pair gE gene.

To confirm the expression of the FIV gag (p50) gene product, cells were infected with S-FHV-016 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal cat anti-FIV (PPR strain) sera was used to detect expression of FIV gag protein. The lysate from S-FHV-016 infected cells exhibited a band at 50 kd which is the expected size of the unprocessed form of the FIV gag protein and a fainter 32 to 35 kd band which may be the result of aberrant cTeavage of the gag protein. In addition, monoclonal antibodies to FIV capsid reacted with a 50 kd protein (the expected size of the FIV gag protein) on a Western blot.

Example 9B S-FHV-017

S-FHV-017 is a recombinant feline herpesvirus that has a deletion of the gE gene and an insertion of two foreign genes at the gE deletion site. The FIV envelope

(env) gene is under the transcriptional control of the

HCMV immediate early promoter and the E. coli lacZ (β- galactosidase) gene is under the transcriptional control of the PRV gX promoter.

S-FHV-017 was derived from S-FHV-001 (NVSL strain) . This was accomplished utilizing the homology vector 725-26.A10 (see Materials and Methods) and virus S- FHV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT HERPESVIRUS. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S-FHV-017. This virus was characterized by restriction endonuclease mapping and the SOUTHERN BLOTTING DNA procedure. This analysis confirmed the insertion of the FIV env gene and the E. coli lacZ (_/β-galactosidase) gene and the deletion of the 1638 base pair gE gene.

To confirm the expression of the FIV env gene product, cells were infected with S-FHV-017 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal cat anti-FIV (PPR strain) sera (J. Elder) was used to detect expression of FIV env protein. The lysate from S-FHV-017 infected cells exhibited a band at 160 kd. This band on a Western corresponds in size to the uncleaved form (SU+TM) of FIV envelope protein. In addition, monoclonal antibodies to gp40 transmembrane domain of FIV env reacts with a band at 160 kd on Western blots.

Example 10:

Recombinant feline herpesvirus expressing antigens from disease causing microorganisms. In addition to recombinant feline herpesvirus expressing antigens from feline leukemia virus, feline immunodeficiency virus and Dirofilaria immi tis (heartworm) . Aadditional antigens from disease causing microorganisms in cats include, but art not limited to Dirofilaria immi tis p39 and 22kD antigens, feline infectious peritonitis virus, calicivirus, rabies, virus, feline parvovirus (panleukopenia virus) , feline coronavirus and feline Chla ydia, Toxoplasma gondii. Disease causing microorganisms in dogs include, but are not limited to canine distemper, canine adenovirus type 1 (hepatitis) , adenovirus type 2 (respiratory disease) , parainfluenza, leptospira canicola, icterohemorragia, parvovirus, coronavirus, borrelia burgdorferi, canine herpesvirus, bordetella bronchiseptica and rabies virus.

Recombinant feline herpesviruses are useful for expressing antigens from disease causing microorganisms from animals in addition to dogs and cats. Recombinant feline herpesvirus is useful as a vaccine in animals including but not limited to humans, horses, cattle, swine and poultry.

Recombinant feline herpesvirus is useful as a vaccine against equine diseases when foreign antigens from the following diseases or disease organisms are expressed in the feline herpesvirus vector: equine influenza, equine herpesvirus-1 and equine herpesvirus-4. Recombinant feline herpesvirus is useful as a vaccine against bovine diseases when foreign antigens from the following diseases, or disease organisms are expressed in the feline herpesvirus vector: bovine viral diarrhea virus, bovine respiratory syncytial virus, bovine parainfluenza virus. Recombinant feline herpesvirus is useful as a vaccine against swine diseases when foreign antigens from the following diseases or disease organisms are expressed in the feline herpesvirus vector: pseudorabies virus, porcine reproductive and respiratory syndrome (PRRS/SIRS) , hog cholera virus, swine influenza virus, swine parvovirus, swine rotavirus. Recombinant feline herpesvirus is useful as a vaccine against poultry diseases when foreign antigens from the following diseases or disease organisms are expressed inthe feline herpesvirus vector: infectious bronchitis virus, Newcastle disease virus, infectious bursal disease virus, Marek's disease virus, infectious laryngotracheitis virus.

Recombinant feline herpesvirus is useful as a vaccine against human diseases. For example, human influenza is a rapidly evolving virus whose neutralizing viral epitopes are rapidly changing. A useful recombinant feline herpesvirus vaccine is one in which the influenza neutralizing eptiopes are quickly changed to protect against new strains of influenza. Human influenza HA and NA genes are cloned into the recombinant feline herpesvirus. Recombinant feline herpesvirus is useful as a vaccine against other human diseases when foreign antigens from the following diseases or disease organisms are expressed in the feline herpesvirus vector: hepatitis B virus surface and core antigens, hepatitis C virus, herpes simplex virus, human herpesviruses, herpes simplex virus-1, herpes simplex virus-2, human herpesvirus-6, human herpesvirus-7, human cytomegalovirus, Epstein-Barr virus, Varicella-Zoster virus, human immunodeficiency virus, human influenza, measles virus, hantaan virus, pneumonia virus, rhinovirus, poliovirus, human respiratory syncytial virus, retrovirus, human T-cell leukemia virus, rabies virus, mumps virus, malaria

( Plasmodium falciparum) , Bordetella pertussis ,

Diptheria, RicJettsia prowazekii , Borrelia bergdorferi , Tetanus toxoid, malignant tumor antigens.

Recombinant feline herpesviruses coexpressing a species-specific cytokine and an antigen from a disease causing microorganism are useful to stimulate an increased cell-mediated and humoral immune response in the animal and increases the efficacy of the recombinant feline herpesvirus as a vaccine. Cytokines which are expressed in feline herpesvirus include but are not limited to interleukin-2, - interleukin-6, interleukin-12, interferon and granulocyte-macrophage colony stimulating factor.

REFERENCES:

1. R. W. Price and A. Kahn, Infection and Immuni ty, 34, 571-580 (1981) .

2. R. B. Tenser, et al . , Journal of Clinical Microbiology 17, 122-127 (1983) .

3. B. Roizman, et al . , Cold Spring Harbor Conference on New Approaches to Viral Vaccines (September

1983) .

K. Fukuchi et al . , Proc . Na tl . Acad. Sci . U. S .A . 82, 751-754 (1985) .

J. M. Koomey et al . , Journal of Virology 50, 662- 665 (1984) .

6. B. Lomniczi et al . , Journal of Virology 49, 970- 979 (1984) .

7. R.A. Crandell and F.D. Maurer, Proc . Soc . Exp. Biol . Med . 97, 487-490 (1958) . 8. R.M. Gaskell and R.C. Povey, Res . Vet . Sci . 27 167-174 (1978) .

9. R.C. Povey, Microbiol. Infect . Dis . 2 373-387

(1979) .

10. K. Tham and M. Studdert, Veterinary Record 120 321-326.

11. F. Fenner, . P.A. Bach ann, E.P.J. Gibbs, F.A. Murphy, M.J. Studdert and D.O. White, Veterinary

Virology, Academic Press, Inc., (1987) .

12. A. Grail, D.A. Harbour, W. Chia, Arch . Virology 116, 209-220 (1991) .

13. P.A. Rota, R.K. Maes and W.T. Ruyechan, Virology 154, 168-179 (1986) .

14. J.H. Nunberg, et al. , J. Virology 63, 3240-3249 (1989) .

15. Federal Register, Vol. 55, No. 90, pp. 19245-19253

16. R. W. Honess, Journal of General Virology 65, 2077-2107 (1984) .

17. M. L. Cook & J. G. Stevens, Journal of General Virology 31, 75-80 (1976) . 18. S. Joshi, et al . , Journal of Virology 65, 5524- 5530 (1991 ) .

19. M. Wachsman, et al . , Journal of General Virology 70, 2513-2520 (1989) .

20. R. A. Bhat, et al . , Nucleic Acids Research 17, 1159-1176 (1989)

21. J. H. Elder and J. I. Mullins, Journal of Virology 46 871-880 (1983) .

22. T. Maniatis, et al . , Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, N.Y.

(1982) .

23. J. Sambrook, et al . , Molecular Cloning: A Laboratory Manual Second Edi tion, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989) . 24. M.A. Innis et al . PCR Protocols : A Guide to Methods and Applications , Academic Press, Inc., San Diego (1990) .

25. Graham and Van der eb (1973), Virology 52, 556-557 (1973) .

26. M. Zijil, et al . , Journal of Virology 62, 2191- 2195 (1988) . 27. M. A. Wild, et al. , 15th International Herpesvirus Workshop, Abstract No. 122, Washington, D.C. (1990) .

28. F. A. Ferrari, et al . , Journal of Bacteriology 161, 556-562 (1985) .

29. B. Lomniczi et al . , Journal of Virology 49 970-979

(1984) . 30. D.J. McGeoch et al . , Journal of Molecular Biology 181 1-13 (1985) .

31. D.R. Thomsen et al . , Gene 16 207-217 (1981) . 32. T.R. Phillips, et al . , J. Virology 64 4605-4613 (1990) .

33. J.P. Ryan, et al . , J. Virology 61 2962-2972 (1987) .

34. J.A. Culpepper, et al . , Mol . Biochem. Parasi tol . 54 51-62 (1992) .

35. J.E. Coligan, ed. , Current Protocols in Immunology John Wiley & Sons, Inc. (1994) 36. G.C. Cole, et al. , J^". Virology 64 4930-4938

(1990) .

37. R.L. Burke, Seminars in Virology 187-197 (1993) .

38. T.C. Mettenleiter, Compara tive Immunology, Mi crobiology and Infectious Diseases 14 151-163

(1991) . 39. U.K. Laemnli, Nature 227, 680-685 (1970)

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANTS:Cochran, Mark D.

McDonell, Michael W.

(ii) TITLE OF INVENTION: Recombinant Feline Herpes virus

(iii) NUMBER OF SEQUENCES: 73

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: John P. White

(B) STREET: 1185 Avenue of the Americas

(C) CITY: New York

(D) STATE: New York

(E) COUNTRY: USA

(F) ZIP: 10036

(v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM 330 466 DX2

(C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: Patentln Release #1.0, Version #1.25

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER: Not Yet Known

(B) FILING DATE: 26-0CT-1995

(C) CLASSIFICATION:

(vii) PRIOR APPLICATION DATA:

(A) APPLICATION NUMBER: 08/329,883

(B) FILING DATE: 26-0CT-1994

(C) CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: White, John P.

(B) REGISTRATION NO: 28,678

(C) REFERENCE/DOCKET NUMBER: 39118-PCT

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: (212)278-0400

(B) TELEFAX: (212)391-0525

(2) INFORMATION FOR SEQ ID NO:1:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2020 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANT -SENSE: NO (Vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 413..2011 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1 :

GGATCCATCA TCAATAGTGC AATACGCAAG CATATAATGG TCTGTGCTGG GCGGCGGATC 60

TATATACCAA ACAACGATGG GCGACCATCA ACGGAAATGA CACGGTTTAC TCGCCAGACT 120

AAACCATCGA ATTCGAGTTC CAAGTCCCTA CTCGATGTCC CCAGATCTTC GAATTCCACC 180

CCAACCGATG GCGTCTCTAG AAGTCAGTTA ACCGTAATTA ACGAAGAAAC CTAATATATT 240

TATAAACAAA TAAAATACTT TTAAAAATGG ATATCTGGTC ATGTGTAATG TTGACGCATA 300

GTGGGTGGTG ACCTAAGATT ATATTAAAAT GTAGAAGGTT TTATGCCCAG TTCACAGTAT 360

CTACTGTGAC CTACCCCGGG GTGGTAATAA CAATACTATC GAATAGCCAA CA ATG 415

Met

1

GGA CTG CTT GTT ACC ATC CTC GTG ATA TTA TTG ATT GTT ACT TCA TCA 463 Gly Leu Leu Val Thr Ile Leu Val Ile Leu Leu Ile Val Thr Ser Ser 5 10 15

AGT TCT ACT ATT CAT CAA GTA ACG ATG ACA GAA GGT GCC GCA CTT TTA 511 Ser Ser Thr Ile His Gin Val Thr Met Thr Glu Gly Ala Ala Leu Leu 20 25 30

GTC GAT GGG GAT GGG ATC GAC CCA CCT TTA AAC AAA ACT TCA CAT TTT 559 Val Asp Gly Asp Gly Ile Asp Pro Pro Leu Asn Lys Thr Ser His Phe 35 40 45

TTG CGA GGT TGG ACA TTT CTA GAG ACT CCG AAA GGA TGT ACA GGA GAG 607 Leu Arg Gly Trp Thr Phe Leu Glu Thr Pro Lys Gly Cys Thr Gly Glu 50 55 60 65

GTG AGT GTT CTA AAA GTA TGT ATA GAT CGT GGG GTA TGT CCG GAT GAT 655 Val Ser Val Leu Lys Val Cys Ile Asp Arg Gly Val Cys Pro Asp Asp 70 75 80

ATC GTT ATA AAT AAG AGA TGT GGT CAC AAA ATG CTT GAA ACC CCA CTA 703 Ile Val Ile Asn Lys Arg Cys Gly His Lys Met Leu Glu Thr Pro Leu 85 90 95

GCG TTG GCG GAA TTT GGA ATT TCT AAT AGT TCT CTC ATC AGA ACC AAA 751 Ala Leu Ala Glu Phe Gly Ile Ser Asn Ser Ser Leu Ile Arg Thr Lys 100 105 110

GAC GTA TAT TTC GTG AAT AAG ACC GTG TTT CCA ATT CTC ACA CCC GAA 799 Asp Val Tyr Phe Val Asn Lys Thr Val Phe Pro Ile Leu Thr Pro Glu 115 120 125

AAA AGT GGC CTT GGT ATT CAG GGG GCC ACT ACG AAT ATA TCC GGG ATA 847 Lys Ser Gly Leu Gly Ile Gin Gly Ala Thr Thr Asn Ile Ser Gly Ile 130 135 140 145

TAT ACC CTG CAT GAG CAC GGT GAT AAT GGA TGG AGT CAT CAA TCT ACA 895 Tyr Thr Leu His Glu His Gly Asp Asn Gly Trp Ser His Gin Ser Thr 150 155 160

TTT TTT GTG ACC GTA AAG GCA AAA CAT CCC GGA CCA TCG TTA ACC CCA 943 Phe Phe Val Thr Val Lys Ala Lys His Pro Gly Pro Ser Leu Thr Pro 165 170 175

GCA CCG GTT CAC TTA ATA ACA CCA CAT CGC CAT GGG GCA CAT TTC CAC 991 Ala Pro Val His Leu Ile Thr Pro His Arg His Gly Ala His Phe His 180 185 190

GTA AGA AAC TAT CAT TCG CAT GTC TAC ATT CCG GGA GAT AAG TTC TTA 1039 Val Arg Asn Tyr His Ser His Val Tyr Ile Pro Gly Asp Lys Phe Leu 195 200 205

TTA GAA ATG CAC CTC AAA TCA GAT ATC TAT GAT CCA GAA TTT TCA GCA 1087 Leu Glu Met His Leu Lys Ser Asp Ile Tyr Asp Pro Glu Phe Ser Ala 210 215 220 225

ACA ATA GAC TGG TAT TTT ATG GAG ACT GAT ATA AAA TGC CCA GTT TTT 1135 Thr Ile Asp Trp Tyr Phe Met Glu Thr Asp Ile Lys Cys Pro Val Phe 230 235 240

AGA ATT TAT GAA ACT TGT ATA TTT CAC CCC CAT GCC GCA TCC TGT CTA 1183 Arg Ile Tyr Glu Thr Cys Ile Phe His Pro His Ala Ala Ser Cys Leu 245 250 255

CAT CCG GAA GAT CCC TCA TGC AGT TTT ACA TCA CCA CTT CGA GCG GTA 1231 His Pro Glu Asp Pro Ser Cys Ser Phe Thr Ser Pro Leu Arg Ala Val 260 265 270

TCT TTA ATT AAT AGA TTT TAT CCA AAA TGC GAT CAC AGA TAT GCC GAT 1279 Ser Leu lie Asn Arg Phe Tyr Pro Lys Cys Asp His Arg Tyr Ala Asp 275 280 285

TGG ACA TCC AGA TGT ATC AAC ACT CCA AGT ATA AAT CAT ATG CCA TAT 1327 Trp Thr Ser Arg Cys Ile Asn Thr Pro Ser Ile Asn His Met Pro Tyr 290 295 300 305

ATC GAA CAG CCG GCC AAT AAC GTG GAT CTA AAG TTT ATC AAT GTA CCC 1375 Ile Glu Gin Pro Ala Asn Asn Val Asp Leu Lys Phe Ile Asn Val Pro 310 315 320

ACC AAC GCT TCT GGG TTG TAC GTA TTC ATA CTT CGT TAT AAT GGA CAT 1423 Thr Asn Ala Ser Gly Leu Tyr Val Phe Ile Leu Arg Tyr Asn Gly His 325 330 335

CCG GAA GAA TGG ACC TAT ACA CTC ATA TCA ACA GGA GCT AAA TTT TTG 1471 Pro Glu Glu Trp Thr Tyr Thr Leu Ile Ser Thr Gly Ala Lys Phe Leu 340 345 350

AAT GTG ATT AGG GAT CTG ACA CGC CCA CGT CTT GGT AGT CAT CAA ATA 1519 Asn Val Ile Arg Asp Leu Thr Arg Pro Arg Leu Gly Ser His Gin Ile 355 360 365

GAG ACC GAT ATT AGC ACA TCT TCG CAG TCG CCT ACC ACG GAG ACA CCA 1567 Glu Thr Asp Ile Ser Thr Ser Ser Gin Ser Pro Thr Thr Glu Thr Pro 370 375 380 385

CGA AAC ATA CAT ATA ACG TGG GCG AGA CGT TAT CTA AAG GTT ATC ATA 1615 Arg Asn Ile His Ile Thr Trp Ala Arg Arg Tyr Leu Lys Val Ile Ile 390 395 400

GGA ATA ATT TGC GTA GCT GGT ATC CTT TTG ATT GTA ATC TCT ATC ACA 1663 Gly Ile Ile Cys Val Ala Gly Ile Leu Leu Ile Val Ile Ser Ile Thr 405 410 415

TGT TAT ATT CGA TTT CGT CAT ATG CGA TAT AAA CCA TAT GAA GTG ATC 1711 Cys Tyr Ile Arg Phe Arg His Met Arg Tyr Lys Pro Tyr Glu Val Ile 420 425 430

AAC CCA TTC CCT GCG GTA TAT ACC AGC ATT CCT AGT AAC GAT CCC GAC 1759 Asn Pro Phe Pro Ala Val Tyr Thr Ser Ile Pro Ser Asn Asp Pro Asp 435 440 445

GAA CTC TAC TTT GAA CGT ATC GCA TCG AAC GAC GAA GAA TCG GCA GAT 1807 Glu Leu Tyr Phe Glu Arg Ile Ala Ser Asn Asp Glu Glu Ser Ala Asp 450 455 460 465 GAT TCT TTT GAT GAA TCA GAT GAG GAG GAG CCA TTG AAT AAT CAT CAT 1855 Asp Ser Phe Asp Glu Ser Asp Glu Glu Glu Pro Leu Asn Asn His His 470 475 480

ATT TCA ACA ACC CAA CAT ACT GAT ATT AAT CCA GAA AAA TCC GGA TCT 1903 Ile Ser Thr Thr Gin His Thr Asp Ile Asn Pro Glu Lys Ser Gly Ser 485 490 495

GGG TAC AGT GTA TGG TTT CGT GAT ACA GAA GAT ACA TCA_CCT CAG CCC 1951 Gly Tyr Ser Val Trp Phe Arg Asp Thr Glu Asp Thr Ser Pro Gin Pro 500 505 510

CTA CAC GCT CCT CCA GAT TAC AGT CGC GTA GTT AAA AGA TTA AAG TCT 1999 Leu His Ala Pro Pro Asp Tyr Ser Arg Val Val Lys Arg Leu Lys Ser 515 520 525

ATT TTA AAA TGACCCGTCG AC 2020

Ile Leu Lys

530

(2) INFORMATION FOR SEQ ID NO:2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 532 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2 :

Met Gly Leu Leu Val Thr Ile Leu Val Ile Leu Leu Ile Val Thr Ser 1 5 10 15

Ser Ser Ser Thr Ile His Gin Val Thr Met Thr Glu Gly Ala Ala Leu 20 25 30

Leu Val Asp Gly Asp Gly Ile Asp Pro Pro Leu Asn Lys Thr Ser His 35 40 45

Phe Leu Arg Gly Trp Thr Phe Leu Glu Thr Pro Lys Gly Cys Thr Gly 50 55 60

Glu Val Ser Val Leu Lys Val Cys Ile Asp Arg Gly Val Cys Pro Asp 65 70 75 80

Asp Ile Val Ile Asn Lys Arg Cys Gly His Lys Met Leu Glu Thr Pro 85 90 95

Leu Ala Leu Ala Glu Phe Gly Ile Ser Asn Ser Ser Leu Ile Arg Thr 100 105 110

Lys Asp Val Tyr Phe Val Asn Lys Thr Val Phe Pro Ile Leu Thr Pro 115 120 125

Glu Lys Ser Gly Leu Gly Ile Gin Gly Ala Thr Thr Asn Ile Ser Gly 130 135 140

Ile Tyr Thr Leu His Glu His Gly Asp Asn Gly Trp Ser His Gin Ser 145 150 155 160

Thr Phe Phe Val Thr Val Lys Ala Lys His Pro Gly Pro Ser Leu Thr 165 170 175

Pro Ala Pro Val His Leu Ile Thr Pro His Arg His Gly Ala His Phe 180 185 190 His Val Arg Asn Tyr His Ser His Val Tyr Ile Pro Gly Asp Lys Phe 195 200 205

Leu Leu Glu Met His Leu Lys Ser Asp Ile Tyr Asp Pro Glu Phe Ser 210 215 220

Ala Thr Ile Asp Trp Tyr Phe Met Glu Thr Asp Ile Lys Cys Pro Val 225 230 235 240

Phe Arg Ile Tyr Glu Thr Cys Ile Phe His Pro His Ala Ala Ser Cys 245 250 255

Leu His Pro Glu Asp Pro Ser Cys Ser Phe Thr Ser Pro Leu Arg Ala 260 265 270

Val Ser Leu Ile Asn Arg Phe Tyr Pro Lys Cys Asp His Arg Tyr Ala 275 280 285

Asp Trp Thr Ser Arg Cys Ile Asn Thr Pro Ser Ile Asn His Met Pro 290 295 300

Tyr Ile Glu Gin Pro Ala Asn Asn Val Asp Leu Lys Phe Ile Asn Val 305 310 315 320

Pro Thr Asn Ala Ser Gly Leu Tyr Val Phe Ile Leu Arg Tyr Asn Gly 325 330 335

His Pro Glu Glu Trp Thr Tyr Thr Leu Ile Ser Thr Gly Ala Lys Phe 340 345 350

Leu Asn Val Ile Arg Asp Leu Thr Arg Pro Arg Leu Gly Ser His Gin 355 360 365

Ile Glu Thr Asp Ile Ser Thr Ser Ser Gin Ser Pro Thr Thr Glu Thr 370 375 380

Pro Arg Asn Ile His Ile Thr Trp Ala Arg Arg Tyr Leu Lys Val Ile 385 390 395 400

Ile Gly Ile Ile Cys Val Ala Gly Ile Leu Leu Ile Val Ile Ser Ile 405 410 415

Thr Cys Tyr Ile Arg Phe Arg His Met Arg Tyr Lys Pro Tyr Glu Val 420 425 430 lie Asn Pro Phe Pro Ala Val Tyr Thr Ser Ile Pro Ser Asn Asp Pro 435 ^~ 440 445

Asp Glu Leu Tyr Phe Glu Arg Ile Ala Ser Asn Asp Glu Glu Ser Ala 450 455 460

Asp Asp Ser Phe Asp Glu Ser Asp Glu Glu Glu Pro Leu Asn Asn His 465 470 475 480

His Ile Ser Thr Thr Gin His Thr Asp Ile Asn Pro Glu Lys Ser Gly 485 490 495

Ser Gly Tyr Ser Val Trp Phe Arg Asp Thr Glu Asp Thr Ser Pro Gin 500 505 510

Pro Leu His Ala Pro Pro Asp Tyr Ser Arg Val Val Lys Arg Leu Lys 515 520 525

Ser Ile Leu Lys 530 (2) INFORMATION FOR SEQ ID NO:3:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline immunodeficiency virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: GCGAGATCTG AAAATGGCCA TTAAGAGATG 30

(2) INFORMATION FOR SEQ ID NO:4:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline immunodeficiency virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4 : GTGCTGCAGT AAAATAGGG 19

(2) INFORMATION FOR SEQ ID NO: 5:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 31 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline immunodeficiency virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: GCCCGGATCC TATGGCAGAA GGGTTTGCAG C 31 (2) INFORMATION FOR SEQ ID NO:6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 31 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline immunodeficiency virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:6 : CCGTGGATCC GGCACTCCAT CATTCCTCCT C 31

(2) INFORMATION FOR SEQ ID NO:7 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline Leukemia Virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: GATCCAATGG GCCCGAACCT CGTCCTGCCT GACCAGAAGC CCCCGTCGAA A 51

(2) INFORMATION FOR SEQ ID NO:8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 14 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: Protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:

Met Gly Pro Asn Leu Val Leu Pro Asp Gin Lys Pro Pro Ser 1 5 - 10

(2) INFORMATION FOR SEQ ID NO:9:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nuclefc acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline Leukemia Virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: GATCCAGCCA AGCTCCGCGA GCGCCTGAAG CAGCGCCAGC AGCTGTTCAA A 51

(2) INFORMATION FOR SEQ ID NO:10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 14 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: Protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:

Ala Lys Leu Arg Glu Arg Leu Lys Glu Arg Glu Gin Leu Phe 1 5 10

(2) INFORMATION FOR SEQ ID NO:11:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus; Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: TACTGTGCAC CTACCCGGGG ATCCTCTAGA GTCGACCTGC AGTGAATAAT AAAATGTGTG 60 TTT 63

(2) INFORMATION FOR SEQ ID NO:12:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 45 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: _ CTCCATAGAA GACACCGGGA CCATGGATCC CGTCGTTTTA CAACG 45

(2) INFORMATION FOR SEQ ID NO:13:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 84 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Feline leukemia virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: GCGGAATTAC AGCTGAGCGC CGGTCGCTAC CATTACCAGT TGGTCTGGTG TCAAAAAGAT 60 CCAATGGGCC CGAACCTCGT CCTG 84

(2) INFORMATION FOR SEQ ID NO:14 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 57 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline leukemia virus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: CAGCAGCTGT TCAAAGATCT AGAATAAGCT AGAGGATCGA TCCCCTATGG CGATCAT 57

(2) INFORMATION FOR SEQ ID NO:15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: CAGTACCGGC GCCTGGTGTC CGTCGACTCT AGAGTCGACC TGCAGGTCGA CGAGTTCTAG CAC 63

(2) INFORMATION FOR SEQ ID NO:16:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 48 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: GGAATTCGAG CTCGGTACCG AGCTCGAATT CCGTGTATTC TATAGTGT 48

(2) INFORMATION FOR SEQ ID NO:17:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 126 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:

AGGTTTTATG CCCAGTTCAC AGTATCTACT GTGCACCTAC CCCGGGGATC CTCTAGAGTC 60

GACCTGCAGC CCAAGCTTGG GCTGCAGGTC GACGAGTTCT AGCACCACGG GAATTGGAAG 120

CTGCTC 126

(2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 33 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: CAGCTGCATT AATGAATCGG CCAACGCGCG GGG 33

(2) INFORMATION FOR SEQ ID NO:19:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 106 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: TTCACAGTAT CTACTGTGCA CCTACCCCGG GGATCCTCTA GAGTCGACCT GCAGCCCAAG 60 CTAGCTTGGC CTCGAGGCCG CGGCCGCCTG CAGGTCGACG TCTGGG 106

(2) INFORMATION FOR SEQ ID NO:20:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: CTCCACAGCT CAACAATGAA GTGGGCAACG TGGATCGATC CCGTCGTTTT ACAACGTCGT 60 GAC 63

(2) INFORMATION FOR SEQ ID NO:21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 48 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: GGGCAGCGTT GGGTCCTGGG ACTCTAGAGG ATCCCCTATG GCGATCAT 48

(2) INFORMATION FOR SEQ ID NO:22: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 111 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:

CAGTACCGGC GCCTGGTGTC CGTCGACCTG CAGGCGGCCG CGGCCTCGAG GCCAAGCTAG 60

CTTGGGCTGC AGGTCGACGA GTTCTAGCAC CACGGGAATT GGAAGCTGCT C 111

(2) INFORMATION FOR SEQ ID NO:23:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: TTCACAGTAT CTACTGTGCA CCTACCCCGG GGATCCTCTA GAGTCGACCT GCAGCCCAAG 60 CTAGCTTGGC CTCGAGGCCA AGCTAGCTTG GGCTGCAGGT CGACGAGTTC TAGCAC 116

(2) INFORMATION FOR SEQ ID NO:24:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus; Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: TACTGTGCAC CTACCCGGGG ATCCTCTAGA GTCGACCTGC AGTGAATAAT AAAATGTGTG 60 TTT 63

(2) INFORMATION FOR SEQ ID NO:25:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 45 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: CTCCATAGAA GACACCGGGA CCATGGCCTC GCTCGCGCGT GCGAT 45

(2) INFORMATION FOR SEQ ID NO:26:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 36 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHET CAL: NO ( iv) ANTI -SENSE : NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Feline leukemia virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: CCGGGCCTCG CGGCCACAGA TCCAATGGGC CCGAAC 36

(2) INFORMATION FOR SEQ ID NO:27:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 45 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline leukemia virus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: CAGAAGCCCC CGTCGAAAGA TCTGAATTCA CTCGAGGGCC TGCCC 45

(2) INFORMATION FOR SEQ ID NO:28:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 55 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: GATCGCCATG GCCAGGATCC TGGCCATCGT GCTGGTCATC CATATGGCGA TCATC 55

(2) INFORMATION FOR SEQ ID NO:29:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: ACGTGGCCTG GTACCGAGCT CGAATTCGAG CTCGCCCGGG GATCCTCTAG AGTCGACGTC 60 TGG 63

(2) INFORMATION FOR SEQ ID NO:30:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 60 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:

CACAGCTCAA CAATGAAGTG GGCAACGTGG ATCGATCCCG TCGTTTTACA ACGTCGTGAC 60

(2) INFORMATION FOR SEQ ID NO: 31:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 60 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Herpes simplex virus type 1

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: GGGCAGCGTT GGGTCCTGGG ACTCTAGAGG ATCCCCGGGA GATGGGGGAG GCTAACTGAA 60

(2) INFORMATION FOR SEQ ID NO:32:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 62 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Herpes simplex virus type 1; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: TGTTGCTGCG TTCCCGGGCG AGCTCGAATT CGAGCTCGGT ACCGCATCGC GGACGGGTGC 60 GC 62

(2) INFORMATION FOR SEQ ID NO:33:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 39 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: CAGTACCGGC GCCTGGTGTC CGTCGACGAG TTCTAGCAC 39

(2) INFORMATION FOR SEQ ID NO:34:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus; Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: TACTGTGCAC CTACCCGGGG ATCCTCTAGA GTCGACCTGC AGTGAATAAT AAAATGTGTG 60 TTT 63 (2) INFORMATION FOR SΕQ ID NO:35:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 46 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus; Feline leukemia virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:35: CTCCATAGAA GACACCGGGA CCATGGATCC AATGGGCCCG AACCTC 46

(2) INFORMATION FOR SEQ ID NO:36:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 42 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline leukemia virus; Hepatitis B virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: GCCTGACCAG AAGCCCCCGT CGAAAGATCC CCGGTGGCTT TG 42

(2) INFORMATION FOR SEQ ID NO:37:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Hepatitis B virus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: CAATGTTAGT ATTCGGGTAC GATCTAGAAT AAGCTAGAGG ATCGATCCCC TATGGCGATC 60 ATC 63

(2) INFORMATION FOR SEQ ID NO:38: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: ACGTGGCCTG GTACCGAGCT CGAATTCGAG CTCGCCCGGG GATCCTCTAG AGTCGACGTC 60 TGG 63

(2) INFORMATION FOR SEQ ID NO:39:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: CTCCACAGCT CAACAATGAA GTGGGCAACG TGGATCGATC CCGTCGTTTT ACAACGTCGT 60 GAC 63

(2) INFORMATION FOR SEQ ID NO:40: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 60 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Herpes simplex virus type 1 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40: GGGCAGCGTT GGGTCCTGGG ACTCTAGAGG ATCCCCGGGA GATGGGGGAG GCTAACTGAA 60

(2) INFORMATION FOR SEQ ID NO:41:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 62 base pairs —

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Herpes simplex virus type 1; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: TGTTGCTGCG TTCCCGGGCG AGCTCGAATT CGAGCTCGGT ACCGCATCGC GGACGGGTGC 60 GC 62

(2) INFORMATION FOR SEQ ID NO:42:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: CAGTACCGGC GCCTGGTGTC CGTCGACTCT AGAGTCGACC TGCAGGTCGA CGAGTTCTAG 60 CAC 63

(2) INFORMATION FOR SEQ ID NO:43:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 99 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO ( iv) ANTI - SENSE : NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus,- Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: TCTACTGTGC ACCTACCCCG GGGATCCTCT AGAGTCGACC TGCAGCCCAA GCTAGCGGCC 60 AATTCAAGCT TGGGCTGCAG TGAATAATAA AATGTGTGT 99

(2) INFORMATION FOR SEQ ID NO:44:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:

GTGAGTTTCT GTGTAACTGA TGATCCTCTA GAGTCGACGT CTGGGGCGCG G 51

(2) INFORMATION FOR SEQ ID NO:45:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:45: CTCCACAGCT CAACAATGAA GTGGGCAACG TGGATCGATC CCGTCGTTTT ACAACGTCGT 60 GAC 63

(2) INFORMATION FOR SEQ ID NO:46:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 95 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS^ double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:46: GCGGAATTAC AGCTGAGCGC CGGTCGCTAC CATTACCAGT TGGTCTGGTG TCAAAAAGAT 60 CTAGAATAAG CTAGAGGATC CCCTATGGCG ATCAT 95

(2) INFORMATION FOR SEQ ID NO:47:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 54 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus,- Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:47: TACCGGCGCC TGGTGTCCGT CGACTCTAGA GGATCATCTG GCGATAGCGC TTAT 54

(2) INFORMATION FOR SEQ ID NO:48:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus; Dirofilaria immitis

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:48: ACCTCCATAG AAGACACCGG GACTCGACTC TAGAGGATCC CCAATTCGGC ACGAGGAAAA 60 AAT ⁶³

(2) INFORMATION FOR SEQ ID NO:49: - 96 -

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO ~

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Dirofilaria immitis; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: AAAAAAAAAA AACTCGAGGG CGAGCTCGAA TTGCTTGGGC TGCAGGTCGA CGAGTTCTAG 60 CAC 63

(2) INFORMATION FOR SEQ ID NO:50:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 48 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:50: GGAATTCGAG CTCGGTACCG AGCTCGAATT CCGTGTATTC TATAGTGT 48

(2) INFORMATION FOR SEQ ID NO:51:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 126 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:51: AGGTTTTATG CCCAGTTCAC AGTATCTACT GTGCACCTAC CCCGGGGATC CTCTAGAGTC 60 GACCTGCAGC CCAAGCTTGG GCTGCAGGTC GACGAGTTCT AGCACCACGG GAATTGGAAG 120 CTGCTC 126

(2) INFORMATION FOR SEQ ID NO:52:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 60 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: CCTTCTGACG GGATTACATC TGCGTTTTTT ACCACAGACA CGGGTACCGA GCTCGAATTC 60

(2) INFORMATION FOR SEQ ID NO:53:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 48 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 53:

GGAATTCGAG CTCGGTACCG AGCTCGAATT CCGTGTATTC TATAGTGT 48

(2) INFORMATION FOR SEQ ID NO:54:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 128 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:

AGGTTTTATG CCCAGTTCAC AGTATCTACT GTGCACCTAC CCCGGGGATC CTCTAGAGTC 60

GACCTGCAGC CCAAGCTAGC GGCCGCTAGC TTGGGCTGCA GGTCGACGAG TTCTAGCACC 120

ACGGGAAT 128

(2) INFORMATION FOR SEQ ID NO:55:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 60 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:55: CCTTCTGACG GGATTACATC TGCGTTTTTT ACCACAGACA CGGGTACCGA GCTCGAATTC 60

(2) INFORMATION FOR SEQ ID NO:56:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 119 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus, Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:56: AGGTTTTATG CCCAGTTCAC AGTATCTACT GTGCACCTAC CCCGGGGATC CTCTAGAGTC 60 GACCTGCAGC CCAAGCTAGC GGCCGCCTGC AGGTCGACGG ACACCAGGCG CCGGTACTG 119

(2) INFORMATION FOR SEQ ID NO:57:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 61 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: CGCACCCGTC CGCGATGCGG TACGGAATTC AAGCTTCTGC AGTGAATAAT AAAATGTGTG 60 T 61

(2) INFORMATION FOR SEQ ID NO:58:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus;

Feline immunodeficiency virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 58: TAGAAGACAC CGGGACTCGA CTCTAGAGGA TCTGAAAATG GCCATTAAGA G 51

(2) INFORMATION FOR SEQ ID NO:59:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 50 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline immunodeficiency virus; Herpes simplex virus type 1

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:59: AGATCCAAAT AATCCATAGA TCCCGGGAGA TGGGGGAGGC TACCTGAAAC 50

(2) INFORMATION FOR SEQ ID NO:60: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 52-baβe pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Herpes simplex virus type 1; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:

TGTTGCGTTC CCCGGGCGAG CTCGAATTCC GTACCAGGCC ACGTGGGCGC GG 52

(2) INFORMATION FOR SEQ ID NO:61:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 99 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: ATGATCGCCA TAGGGGATCG ATCCTCTAGC TTATTCTAGA TCTTTTTGAC ACCAGACCAA 60 CTGGTAATGG TAGCGACCGG CGCTCAGCTG TAATTCCGC 99

(2) INFORMATION FOR SEQ ID NO:62:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:62: CCAGTCACGA CGTTGTAAAA CGACGGGATC GATCCACGTT GCCCACTTCA TTGTTGAGCT 60 GTG 63

(2) INFORMATION FOR SEQ ID NO:63: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:63: CCCAGACGTC GACCTGCAGG CGGCCGCTAG CTTGGGCTGC AGGTCGACGA GTTCTAGCAC 60 CAC 63

(2) INFORMATION FOR SEQ ID NO:64:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 90 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline herpesvirus; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:64: TCTACTGTGC ACCTACCCCG GGGATCCTCT AGAGTCGACC TGCAGCCCAA GCTAGCGGCC 60

GCGTTTAAAC GTCGACGTCT GGGGCGCGGG 90

(2) INFORMATION FOR SEQ ID NO:65:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; E. coli

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:65: CTCCACAGCT CAACAATGAA GTGGGCAACG TGGATCGATC CCGTCGTTTT ACAACGTCGT 60 GAC 63

(2) INFORMATION FOR SEQ ID NO:66:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 95 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: E. coli; Pseudorabies virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:

GCGGAATTAC AGCTGAGCGC CGGTCGCTAC CATTACCAGT TGGTCTGGTG TCAAAAAGAT 60

CTAGAATAAG CTAGAGGATC CCCTATGGCG ATCAT 95

(2) INFORMATION FOR SEQ ID NO:67:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 63 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Pseudorabies virus; Herpes simplex virus type 1

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:67: CTGGTGTCCG TCGAGTCGAG ATCCTCTAGA GTCGACAAGC TTGGGCTGCA GGTCGGGAAC 60 GCA 63

(2) INFORMATION FOR SEQ ID NO:68:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 64 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE: (A) ORGANISM: Herpes simplex virus type 1; Feline immunodeficiency virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: GCCTCCCCCA TCTCCCGACT CTAGCTAGAG GAGGCACTCC ATCATTCCTC CTCTTTTTCA 60 GACA 64

(2) INFORMATION FOR SEQ ID NO:69:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 60 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline immunodeficiency virus; Human cytomegalovirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: CATTGTCTAT TGGCTGCAAA CCCTTCTGCC ATAGGATCCA TGGTCCCGGT GTCTTCTATG 60

(2) INFORMATION FOR SEQ ID NO:70:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 90 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Human cytomegalovirus; Feline herpesvirus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:70: CACACATTTT ATTATTCACT GCAGCCCAAG CTTCCCGGGA GTTTAAACGC GGCCGCTAGC 60 TTGGGCTGCA GGTCGACGAG TTCTAGCACC 90

(2) INFORMATION FOR SEQ ID NO:71:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2330 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE:

(A) ORGANISM: Feline Herpesvirus

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 2..904

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1165..2283

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 71:

C TTC GAA GGT CTA ACC ATC TAT ACC TTG GTA AAT ATA AGT CTA CTC 46

Phe Glu Gly Leu Thr Ile Tyr Thr Leu Val Asn Ile Ser Leu Leu 1 5 10 15

CTT CAA CCA GGA ATA TTC GAT TCC GGG AGT TTC CTG TAT TCA TTT ATA 94 Leu Gin Pro Gly Ile Phe Asp Ser Gly Ser Phe Leu Tyr Ser Phe Ile 20 25 30

TAT GGT CAA AAT AGA TAC AAT GGA CGT ATT ATA GTT CAT GTA GAA AAA 142 Tyr Gly Gin Asn Arg Tyr Asn Gly Arg Ile Ile Val His Val Glu Lys 35 40 45

AAT ACT GAT TAT CCC TGC AAA ATG TAT CAT GGA CTC ATG GCT CCA TTT 190 Asn Thr Asp Tyr Pro Cys Lys Met Tyr His Gly Leu Met Ala Pro Phe 50 55 60

GAC CAT CAT CCC CAA AGC CAC GTT GAA ACT CCG AAT GAT AAG AAT CAT 238 Asp His His Pro Gin Ser His Val Glu Thr Pro Asn Asp Lys Asn His 65 70 75

CGT AGA GGG CGG GGA TGT TTT CCC GAA TTG GTG GAA CCT GTT CTA TGG 286 Arg Arg Gly Arg Gly Cys Phe Pro Glu Leu Val Glu Pro Val Leu Trp 80 85 90 95

GTT AAT ATC AGC AGT GAT CTT ATT GGT GGT CCA CCT TTC GAC TAT AAT 334 Val Asn Ile Ser Ser Asp Leu Ile Gly Gly Pro Pro Phe Asp Tyr Asn 100 105 110

CAT GAA GAT GAG GCT GAT ATT GAG AGT GAT GAG CTC CCG GAG GAG GAT 382 His Glu Asp Glu Ala Asp Ile Glu Ser Asp Glu Leu Pro Glu Glu Asp 115 120 125

ATA CAT AAC TAC TCA GAT TGT CGT GCG ACT AAT ATG TTT GTT CCG AGA 430 Ile His Asn Tyr Ser Asp Cys Arg Ala Thr Asn Met Phe Val Pro Arg 130 135 140

GAG CCC CTC AGT CAA GTT CTT GGT TCT CAA AGT CTA CTG GTT GGT AGT 478 Glu Pro Leu Ser Gin Val Leu Gly Ser Gin Ser Leu Leu Val Gly Ser 145 150 155

TTA GGT TTC CAG ATA ATT ACT CAA CCC TGG CAA CTG AAG CAG AAT GAA 526 Leu Gly Phe Gin Ile Ile Thr Gin Pro Trp Gin Leu Lys Gin Asn Glu 160 165 170 175

AGT TAT GAT GGA CTA AGA AAT GCC TCT CTT GAA CCC CGA CAC CTT GAC 574 Ser Tyr Asp Gly Leu Arg Asn Ala Ser Leu Glu Pro Arg His Leu Asp 180 185 190

TCC AGT AAC GAT CGT GAT CTA CTA GAT GAA ACT GAA ATG ATT GGA TCG 622 Ser Ser Asn Asp Arg Asp Leu Leu Asp Glu Thr Glu Met Ile Gly Ser 195 200 205

ATT ATT ACG ACT CCA CCA CCA ACC CAT CCA AAA GGT GTC AAT GGG GGT 670 Ile Ile Thr Thr Pro Pro Pro Thr His Pro Lys Gly Val Asn Gly Gly 210 215 220

TTC CTC CAA GAT CTA CCA ATT ATC GAG CCT ACG ACC GAA CCA TGC TTA 718 Phe Leu Gin Asp Leu Pro Ile Ile Glu Pro Thr Thr Glu Pro Cys Leu 225 230 235

GTA CAT ACA AAG ATC ATT GGG ATC GGA ACA GTA GTC GTT GTA TTT TTG 766 Val His Thr Lys Ile Ile Gly Ile Gly Thr Val Val Val Val Phe Leu 240 245 250 255

TTA TTT ATT CTC ATA TCC CTA TGT GTT TAT ACT TGC GTT CTA CGA TCC 814 Leu Phe Ile Leu Ile Ser Leu Cys Val Tyr Thr Cys Val Leu Arg Ser 260 265 270

CGC ATC GGT ATG GTA GAT CGC GCC TAT GTG AAA CAA GTA CGA TTT AAT 862 Arg Ile Gly Met Val Asp Arg Ala Tyr Val Lys Gin Val Arg Phe Asn 275 280 285

TCC AAT CCA TCA TAT CAA CAG TTG ACA AGA TAC CCC CAA CCA 904

Ser Asn Pro Ser Tyr Gin Gin Leu Thr Arg Tyr Pro Gin Pro 290 295 300

TAATAAACTG ATTAAATTTA ATTAAAGTCT CATATGTGGG GCTGTGGGGA CGAGGGGCTG 964

TGGGGACGAG GGGCTGTGGG GACGATTACA ACCGATAAAT GTCGTATATG AAATGTGGTG 1024

TTAACATAAC ACGGATTTTT TAAGCACACC ACATGACACA CCCCCACGAT AACGGTTTAA 1084

ATCACCAGCT ATGTGAACTG CCCTCCATTC TACTCAAATG AGTGGTGGTG TGTGGCATAT 1144 TAGAACCATT TCGTCTAATG ATG ACA CGT CTA CAT TTT TGG TGG TGT GGA 1194

Met Thr Arg Leu His Phe Trp Trp Cys Gly 1 5 10

ATC TTT GCG GTC CTG AAA TAT CTG GTA TGT ACT TCA AGC CTT ACG ACC 1242

Ile Phe Ala Val Leu Lys Tyr Leu Val Cys Thr Ser Ser Leu Thr Thr

15 20 25

ACG CCA AAA ACA ACT ACG GTT TAT GTG AAG GGA TTT AAT ATA CCT CCA 1290

Thr Pro Lys Thr Thr Thr Val Tyr Val Lys Gly Phe Asn Ile Pro Pro 30 35 40

CTA CGC TAC AAT TAT ACT CAA GCC AGA ATC GTG CCA AAA ATT CCC CAG 1338 Leu Arg Tyr Asn Tyr Thr Gin Ala Arg Ile Val Pro Lys Ile Pro Gin 45 50 55

GCG ATG GAT CCG AAG ATA ACA GCT GAA GTA CGT TAT GTA ACA TCA ATG 1386 Ala Met Asp Pro Lys Ile Thr Ala Glu Val Arg Tyr Val Thr Ser Met 60 65 70

GAT TCA TGT GGG ATG GTG GCA TTG ATA TCA GAG CCG GAT ATA GAC GCT 1434 Asp Ser Cys Gly Met Val Ala Leu Ile Ser Glu Pro Asp Ile Asp Ala 75 80 85 90

ACT ATT CGA ACC ATA CAA CTA TCT CAA AAA AAA ACA TAT AAC GCG ACT 1482 Thr Ile Arg Thr lie Gin Leu Ser Gin Lys Lys Thr Tyr Asn Ala Thr 95 100 105

ATA AGT TGG TTT AAG GTA ACC CAG GGT TGT GAA TAC CCT ATG TTT CTT 1530 Ile Ser Trp Phe Lys Val Thr Gin Gly Cys Glu Tyr Pro Met Phe Leu 110 115 120

ATG GAT ATG AGA CTT TGT GAT CCT AAA CGG GAA TTT GGA ATA TGT GCT 1578 Met Asp Met Arg Leu Cys Asp Pro Lys Arg Glu Phe Gly Ile Cys Ala 125 130 135

TTA CGG TCG CCT TCA TAT TGG TTG GAA CCT TTA ACA AAG TAT ATG TTC 1626 Leu Arg Ser Pro Ser Tyr Trp Leu Glu Pro Leu Thr Lys Tyr Met Phe 140 145 150

CTA ACA GAC GAT GAA CTG GGT TTG ATT ATG ATG GCC CCG GCC CAA TTT 1674 Leu Thr Asp Asp Glu Leu Gly Leu Ile Met Met Ala Pro^"Ala Gin Phe 155 160 165 170

AAT CAA GGA CAA TAT CGA AGA GTT ATA ACC ATC GAT GGT TCC ATG TTT 1722 Asn Gin Gly Gin Tyr Arg Arg Val Ile Thr Ile Asp Gly Ser Met Phe 175 180 185

TAT ACA GAT TTT ATG GTA CAA CTA TCT CCA ACG CCA TGT TGG TTC GCA 1770 Tyr Thr Asp Phe Met Val Gin Leu Ser Pro Thr Pro Cys Trp Phe Ala 190 195 200

AAA CCC GAT AGA TAC GAA GAG ATT CTA CAT GAA TGG TGT CGA AAT GTT 1818 Lys Pro Asp Arg Tyr Glu Glu Ile Leu His Glu Trp Cys Arg Asn Val 205 210 215

AAA ACT ATT GGC CTT GAT GGA GCT CGT GAT TAC CAC TAT TAT TGG GTA 1866 Lys Thr Ile Gly Leu Asp Gly Ala Arg Asp Tyr His Tyr Tyr Trp Val 220 225 230

CCC TAT AAC CCA CAA CCT CAC CAT AAA GCC GTA CTC TTA TAT TGG TAT 1914 Pro Tyr Asn Pro Gin Pro His His Lys Ala Val Leu Leu Tyr Trp Tyr 235 240 245 250

CGG ACT CAT GGC CGA GAA CCC CCA GTA AGA TTC CAA GAG GCC ATT CGA 1962 Arg Thr His Gly Arg Glu Pro Pro Val Arg Phe Gin Glu Ala Ile Arg 255 260 265

TAT GAT CGT CCC GCC ATA CCG TCT GGG AGT GAG GAT TCG AAA CGG TCC 2010 Tyr Asp Arg Pro Ala Ile Pro Ser Gly Ser Glu Asp Ser Lys Arg Ser 270 275 280

AAC GAC TCT AGA GGA GAA TCG AGT GGA CCC AAT TGG ATA GAC ATT GAA 2058 Asn Asp Ser Arg Gly Glu Ser Ser Gly Pro Asn Trp Ile Asp Ile Glu 285 290 295

AAT TAC ACT CCT AAA AAT AAT GTG CCT ATT ATA ATA TCT GAC GAT GAC 2106 Asn Tyr Thr Pro Lys Asn Asn Val Pro Ile Ile Ile Ser Asp Asp Asp 300 305 310

GTT CCT ACA GCC CCT CCC AAG GGC ATG AAT AAT CAG TCA GTA GTG ATA 2154 Val Pro Thr Ala Pro Pro Lys Gly Met Asn Asn Gin Ser Val Val Ile 315 320 325 330

CCC GCA ATC GTA CTA AGT TGT CTT ATA ATA GCA CTG ATT CTA GGA GTG 2202 Pro Ala Ile Val Leu Ser Cys Leu Ile Ile Ala Leu Ile Leu Gly Val 335 340 345

ATA TAT TAT ATT TTG AGG GTA AAG AGG TCT CGA TCA ACT GCA TAT CAA 2250 Ile Tyr Tyr Ile Leu Arg Val Lys Arg Ser Arg Ser Thr Ala Tyr Gin 350 355 360

CAA CTT CCT ATA ATA CAT ACA ACT CAC CAT CCT TAAGCTCCAC ATTCCAATCG 2303 Gin Leu Pro Ile Ile His Thr Thr His His Pro 365 370

AGTTGGTAGG GAAGATATGA AGTGGGC 2330 (2) INFORMATION FOR SEQ ID NO:72:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 301 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 72:

Phe Glu Gly Leu Thr Ile Tyr Thr Leu Val Asn Ile Ser Leu Leu Leu 1 5 10 15

Gin Pro Gly Ile Phe Asp Ser Gly Ser Phe Leu Tyr Ser Phe Ile Tyr 20 25 30

Gly Gin Asn Arg Tyr Asn Gly Arg Ile Ile Val His Val Glu Lys Asn 35 40 45

Thr Asp Tyr Pro Cys Lys Met Tyr His Gly Leu Met Ala Pro Phe Asp 50 55 60

His His Pro Gin Ser His Val Glu Thr Pro Asn Asp Lys Asn His Arg 65 70 75 80

Arg Gly Arg Gly Cys Phe Pro Glu Leu Val Glu Pro Val Leu Trp Val 85 90 95

Asn Ile Ser Ser Asp Leu Ile Gly Gly Pro Pro Phe Asp Tyr Asn His 100 105 110

Glu Asp Glu Ala Asp Ile Glu Ser Asp Glu Leu Pro Glu Glu Asp Ile 115 120 125

His Asn Tyr Ser Asp Cys Arg Ala Thr Asn Met Phe Val Pro Arg Glu 130 135 140

Pro Leu Ser Gin Val Leu Gly Ser Gin Ser Leu Leu Val Gly Ser Leu 145 150 155 160

Gly Phe Gin Ile Ile Thr Gin Pro Trp Gin Leu Lys Gin Asn Glu Ser 165 170 175

Tyr Asp Gly Leu Arg Asn Ala Ser Leu Glu Pro Arg His Leu Asp Ser 180 185 190

Ser Asn Asp Arg Asp Leu Leu Asp Glu Thr Glu Met Ile Gly Ser Ile 195 200 205

Ile Thr Thr Pro Pro Pro Thr His Pro Lys Gly Val Asn Gly Gly Phe 210 215 220

Leu Gin Asp Leu Pro Ile Ile Glu Pro Thr Thr Glu Pro Cys Leu Val 225 230 235 240

His Thr Lys Ile Ile Gly Ile Gly Thr Val Val Val Val Phe Leu Leu 245 250 255

Phe Ile Leu Ile Ser Leu Cys Val Tyr Thr Cys Val Leu Arg Ser Arg 260 265 270

Ile Gly Met Val Asp Arg Ala Tyr Val Lys Gin Val Arg Phe Asn Ser 275 280 285

Asn Pro Ser Tyr Gin Gin Leu Thr Arg Tyr Pro Gin Pro 290 295 300 (2) INFORMATION FOR SEQ ID NO:73:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 373 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:

Met Thr Arg Leu His Phe Trp Trp Cys Gly Ile Phe Ala Val Leu Lys 1 5 10 15

Tyr Leu Val Cys Thr Ser Ser Leu Thr Thr Thr Pro Lys Thr Thr Thr 20 25 30

Val Tyr Val Lys Gly Phe Asn Ile Pro Pro Leu Arg Tyr Asn Tyr Thr 35 40 45

Gin Ala Arg Ile Val Pro Lys Ile Pro Gin Ala Met Asp Pro Lys Ile 50 55 60

Thr Ala Glu Val Arg Tyr Val Thr Ser Met Asp Ser Cys Gly Met Val 65 70 75 80

Ala Leu Ile Ser Glu Pro Asp Ile Asp Ala Thr Ile Arg Thr Ile Gin 85 90 95

Leu Ser Gin Lys Lys Thr Tyr Asn Ala Thr Ile Ser Trp Phe Lys Val 100 105 110

Thr Gin Gly Cys Glu Tyr Pro Met Phe Leu Met Asp Met Arg Leu Cys 115 120 125

Asp Pro Lys Arg Glu Phe Gly Ile Cys Ala Leu Arg Ser Pro Ser Tyr 130 135 140

Trp Leu Glu Pro Leu Thr Lys Tyr Met Phe Leu Thr Asp Asp Glu Leu 145 150 155 160

Gly Leu Ile Met Met Ala Pro Ala Gin Phe Asn Gin Gly Gin Tyr Arg 165 170 175

Arg Val Ile Thr Ile Asp Gly Ser Met Phe Tyr Thr Asp Phe Met Val 180 185 190

Gin Leu Ser Pro Thr Pro Cys Trp Phe Ala Lys Pro Asp Arg Tyr Glu 195 200 205

Glu Ile Leu His Glu Trp Cys Arg Asn Val Lys Thr Ile Gly Leu Asp 210 215 220

Gly Ala Arg Asp Tyr His Tyr Tyr Trp Val Pro Tyr Asn Pro Gin Pro 225 230 235 240

His His Lys Ala Val Leu Leu Tyr Trp Tyr Arg Thr His Gly Arg Glu 245 250 255

Pro Pro Val Arg Phe Gin Glu Ala Ile Arg Tyr Asp Arg Pro Ala Ile

260 265 270

Pro Ser Gly Ser Glu Asp Ser Lys Arg Ser Asn Asp Ser Arg Gly Glu 275 280 285

Ser Ser Gly Pro Asn Trp Ile Asp Ile Glu Asn Tyr Thr Pro Lys Asn 290 295 300

Asn Val Pro Ile Ile Ile Ser Asp Asp Asp Val Pro Thr Ala Pro Pro 305 310 315 320

Lys Gly Met Asn Asn Gin Ser Val Val Ile Pro Ala Ile Val Leu Ser 325 330 335

Cys Leu Ile Ile Ala Leu Ile Leu Gly Val Ile Tyr Tyr Ile Leu Arg 340 345 350

Val Lys Arg Ser Arg Ser Thr Ala Tyr Gin Gin Leu Pro Ile Ile His 355 360 365

Thr Thr His His Pro 370

Claims

What is claimed is:

1. A recombinant feline herpes virus comprising a feline herpes virus viral genome which contains a deletion in the unique short region of the viral genome, wherein the deletion is in the glycoprotein E (gE) gene.

2. The recombinant feline herpes virus of claim 1, wherein the feline herpes virus further contains a foreign DNA sequence inserted into a non- essential region of the feline herpes virus genome.

3. The recombinant feline herpes virus of claim 2, wherein the foreign DNA sequence is inserted into a unique short region of the feline herpes virus.

4. The recombinant feline herpes virus of claim 3, wherein the foreign DNA sequence is inserted in the deleted gE gene.

5. The recombinant feline herpes virus of claim 1, further characterized by a deletion in a glycoprotein I (gl) gene.

6. The recombinant feline herpes virus of claim 1 further characterized by a deletion in a glycoprotein G (gG) gene.

7. The recombinant feline herpes virus of claim 2, which further comprises the foreign DNA sequence inserted into the unique short region, wherein the foreign gene is capable of being expressed in a recombinant feline herpes virus host cell.

8. The recombinant feline herpes virus of claim 2, wherein the foreign DNA sequence encodes a screeneable marker.

9. The recombinant feline herpes virus of claim 8, wherein the screenable marker is E. coli beta- galactosidase. ^~~

10. The recombinant feline herpes virus of claim 8, wherein the screenable marker is E. coli beta- glucouronidase.

11. The recombinant feline herpes virus of claim 2, wherein the foreign DNA sequence encodes an antigenic polypeptide.

12. The recombinant feline herpes virus of claim 11, wherein the antigenic polypeptide when introduced into the host cell, induces production of protective antibodies against a feline disease causing agent from which the antigenic polypeptide is derived or derivable.

13. The recombinant feline herpes virus of claim 12, wherein the antigenic polypeptide is derived or derivable from a group consisting of feline pathogen, canine pathogen, equine pathogen, bovine pathogen, avian pathogen, porcine pathogen, or human pathogen.

14. The recombinant feline herpes virus of claim 2, wherein the foreign DNA sequence is selected from a group consisting of feline leukemic virus envelope gene, hepatitis B core antigen gene, Pseudorabies virus glycoprotein C gene, Dirofilaria immi tis 22 kD or p39 gene, feline immunodeficiency virus gag, pol, or env genes, or E. coli beta-galactosidase gene.

15. The recombinant feline herpes virus of claim 14, wherein the foreign gene is a fusion protein of two or more foreign genes.

16. The recombinant feline herpes virus of claim 2, wherein the foreign gene is under control of an endogenous upstream feline herpes virus promoter.

17. The recombinant feline herpes virus of claim 2, wherein the foreign gene is under control of a heterologous upstream promoter.

18. The recombinant feline herpes virus of claim 16, wherein promoter is selected from a group consisting of the HCMV IE promoter, PRV gX promoter or BHV-1.1 VP8.

19. The recombinant feline herpes virus of claim 1 designated S-FHV-003.

20. The recombinant feline herpes virus of claim 1 designated S-FHV-004 (ATCC Accession No. VR 2487) .

21. The recombinant feline herpes virus of claim 1 designated S-FHV-006.

22. The recombinant feline herpes virus of claim 1 designated S-FHV-007.

23. The recombinant feline herpes virus of claim 1 designated S-FHV-009.

24. The recombinant feline herpes virus of claim 1 designated S-FHV-012.

25. The recombinant feline herpes virus of claim 1 designated^" S-FHV-008.

26. The recombinant feline herpes virus of claim 1 designated S-FHV-011.

27. The recombinant feline herpes virus of claim 1 designated S-FHV-005.

28. The recombinant feline herpes virus of claim 1 designated S-FHV-010.

29. The recombinant feline herpes virus of claim 1 designated S-FHV-014 (ATCC Accession No. VR 2488) .

30. The recombinant feline herpes virus of claim 1 designated S-FHV-016.

31. The recombinant feline herpes virus of claim 1 designated S-FHV-017.

32. A recombinant feline herpes virus comprising a feline herpes virus viral genome which contains an insertion into the unique short region of the viral genome, wherein the insertion is in the glycoprotein E (gE) gene.

33. The recombinant feline herpes virus of claim 32, wherein the feline herpes virus further contains a deletion in the unique short region of the feline herpes virus genome.

34. The recombinant feline herpes virus of claim 33 further characterized by a deletion in a glycoprotein I (gl) gene.

35. The recombinant feline herpes virus of claim 33 further characterized by a deletion in a glycoprotein G (gG) gene.

36. A homology vector for producing a recombinant feline herpes virus by inserting a foreign DNA sequence into the feline herpes virus genome which comprises a double-stranded DNA molecule consisting of: a) double-stranded foreign DNA sequence encoding an antigenic polypeptide derived from an animal pathogen;

b) at one end of the foreign DNA sequence, double-stranded feline virus genomic DNA homologous to the genomic DNA located at one side of a non-essential site of the feline herpes viral genomic DNA;

c) at the other end of the foreign DNA sequence, double stranded feline herpes virus genomic DNA homologous to the genomic DNA located at the other side of the same site.

37. The homology vector of claim 36, wherein the antigenic polypeptide is selected from a group consisting of hepatitis B core antigen protein, pseudorabies virus glycoprotein C, feline leukemia virus envelope, Dirofilaria immi tis 22 kD or p39 gene, feline immunodeficiency virus gag, pol, or env genes, or E. coli beta-galactosidase gene .

38. The homology vector of claim 36, designated Homology Vector 416-80.2IB.

39. The homology vector of claim 36, designated Homology Vector 644-09.Al.

40. The homology vector of claim 36, designated Homology Vector 644-09.B2.

41. The homology vector of claim 36, designated Homology Vector 478-17. IT.

42. The homology vector of claim 36, designated Homology Vector 478.10.11.

43. The homology vector of claim 36, designated Homology Vector 411-91.01A.

44. The homology vector of claim 36, designated Homology Vector 411-91.01B.

45. The homology vector of claim 36, designated Homology Vector 510-36.1C.

46. The homology vector of claim 36, designated Homology Vector 510-36.2F.

47. The homology vector of claim 36, designated Homology Vector 416-88.2L.

48. The homology vector of claim -36, designated Homology Vector 639.85.1H.

49. The homology vector of claim 36, designated Homology Vector 669-42.04.

50. The homology vector of claim 36, designated Homology Vector 725-26.A10.

51. A homology vector for producing a recombinant feline herpes virus by inserting a foreign DNA sequence into the feline herpes virus genome which comprises a double-stranded DNA molecule consisting of: a) double-stranded foreign DNA sequence encoding an antigenic polypeptide derived from a cytokine capable of stimulating an immune response;

b) at one end of the foreign DNA sequence, double-stranded feline herpes virus genomic DNA homologous to the genomic DNA located at one side of a non- essential site of the feline herpes virus genomic DNA;

c) at the other end of the foreign DNA sequence, double stranded feline virus genomic DNA homologous to the genomic

DNA located at the other side of the same site.

52. A host cell infected with the recombinant feline herpes virus of claim 1.

53. The host cell of claim 52, wherein the host cell is a mammalian cell .

5 . A vaccine against feline herpes virus which comprises an effective immunizing amount of the recombinant feline herpes virus of claim 1 and a suitable carrier.

55. A vaccine against an animal pathogen which comprises an effective immunizing amount of the recombinant feline herpes virus of claim 2 and a suitable carrier.

56. A method of immunizing an animal against an animal pathogen which comprises administering to the animal an effective immunizing dose of the vaccine of claim 55 .

57. A method of immunizing an animal against an feline pathogen which comprises administering to the animal an effective immunizing dose of the vaccine of claim 55.

58. A method of distinguishing an animal vaccinated with the vaccine of claim 1 from an animal infected with a naturally-occuring feline herpes virus which comprises analysing a sample of a body fluid from the animal for the presence of feline herpes virus gE and at least one other antigen normally expressed in an animal infected by a naturally-occuring feline herpes virus, determining whether the antigen and gE are present in the body fluid, the presence of the antigen and the absence of gE indicative of an animal vaccinated with the vaccine and not infected with a naturally-occuring feline herpes virus.

59. The method of claim 58, wherein the presence of the antigen and of gE in the body fluid is determined by detecting in the body fluid antibodies specific for the antigen and for gE.