WO1999049077A1 - Fluorescent assay for topoisomerase inhibitors - Google Patents
Fluorescent assay for topoisomerase inhibitors Download PDFInfo
- Publication number
- WO1999049077A1 WO1999049077A1 PCT/IB1999/000473 IB9900473W WO9949077A1 WO 1999049077 A1 WO1999049077 A1 WO 1999049077A1 IB 9900473 W IB9900473 W IB 9900473W WO 9949077 A1 WO9949077 A1 WO 9949077A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- dna
- fluid
- topoisomerase
- test compound
- gyrase
- Prior art date
Links
- 239000003534 dna topoisomerase inhibitor Substances 0.000 title claims abstract description 6
- 229940044693 topoisomerase inhibitor Drugs 0.000 title claims abstract description 6
- 238000003556 assay Methods 0.000 title description 11
- 239000012530 fluid Substances 0.000 claims abstract description 36
- 150000001875 compounds Chemical class 0.000 claims abstract description 35
- 238000012360 testing method Methods 0.000 claims abstract description 32
- 101710183280 Topoisomerase Proteins 0.000 claims abstract description 29
- 238000000034 method Methods 0.000 claims abstract description 23
- ZYFVNVRFVHJEIU-UHFFFAOYSA-N PicoGreen Chemical compound CN(C)CCCN(CCCN(C)C)C1=CC(=CC2=[N+](C3=CC=CC=C3S2)C)C2=CC=CC=C2N1C1=CC=CC=C1 ZYFVNVRFVHJEIU-UHFFFAOYSA-N 0.000 claims abstract description 9
- 230000000694 effects Effects 0.000 claims abstract description 7
- 239000006184 cosolvent Substances 0.000 claims description 5
- 230000005284 excitation Effects 0.000 claims description 4
- 239000013612 plasmid Substances 0.000 claims description 3
- 241000588724 Escherichia coli Species 0.000 claims description 2
- 108020004707 nucleic acids Proteins 0.000 abstract description 10
- 102000039446 nucleic acids Human genes 0.000 abstract description 10
- 239000000980 acid dye Substances 0.000 abstract description 8
- -1 cyanine nucleic acid Chemical class 0.000 abstract description 7
- 108020004414 DNA Proteins 0.000 description 41
- 239000000975 dye Substances 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 102000003915 DNA Topoisomerases Human genes 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 108090000323 DNA Topoisomerases Proteins 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 239000003068 molecular probe Substances 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 108020000946 Bacterial DNA Proteins 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 2
- 229960005542 ethidium bromide Drugs 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 229940124307 fluoroquinolone Drugs 0.000 description 2
- 238000012188 high-throughput screening assay Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 2
- 239000012536 storage buffer Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 230000007018 DNA scission Effects 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000007421 fluorometric assay Methods 0.000 description 1
- 239000002271 gyrase inhibitor Substances 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000002821 scintillation proximity assay Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000011877 solvent mixture Substances 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/533—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving isomerase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/99—Isomerases (5.)
Definitions
- the subject invention relates to analytical procedures which use fluorescent dyes for detection of and/or for providing a quantitative measure of substances which inhibit topoisomerase enzymes, especially those of bacteria.
- Gyrase a type II topoisomerase
- fluoroquinolones popular commercial antibacterials
- gyrase mediates bacterial DNA supercoiling which is essential for DNA metabolism and bacterial survival and replication.
- the subject invention involves methods for determining the activity of test compounds as topoisomerase inhibitors by incubating a DNA with the topoisomerase of interest, both in the presence and absence of the test compound of interest, incorporating a cyanine nucleic acid dye, and comparing the fluorescence from the dye when the test compound is present and absent.
- inhibitors e.g., fluoroquinolones
- topoisomerases e.g., gyrase
- Dyes such as PicoGreen® differentially bind to relaxed and supercoiled topological isomers of duplex DNA. Unlike other nucleic acid dyes, they reproducibly fluoresce more intensely upon binding to supercoiled DNA (product) following gyrase-mediated supercoiling of relaxed DNA, allowing greater discrimination between DNA topoisomers than achieved by previous assays. Inhibitors of gyrase are identifiable by reduction in fluorescence compared to control.
- the subject invention involves methods for determining activity of test compounds as topoisomerase inhibitors comprising the following steps:
- step (c) incorporating a cyanine nucleic acid dye in the incubated fluid from step (b);
- step (e) repeating steps (a) to (d) omitting the test compound from the fluid of step (a);
- step (f) optionally repeating steps (a) to (d) omitting the test compound and the topoisomerase from the fluid of step (a);
- the liquid component(s) of the fluid are selected such that the test compound, DNA and topoisomerase are intimately mixed, preferably as a fine suspension, or more preferably in solution.
- the fluid is preferably aqueous-based, but may be based on another solvent or a solvent mixture, e.g., a water/cosolvent mixture.
- Cosolvents mixed with water for this purpose include, e.g., dimethyl sulfoxide (DMSO) and N, N-dimethylformamide (DMF).
- DMSO dimethyl sulfoxide
- DMF N, N-dimethylformamide
- the fluid is preferably an aqueous solution or an aqueous/cosolvent solution containing up to about 2% cosolvent.
- more than one concentration of the test compound is tested using a subject invention method, to ascertain the range of concentrations, and particularly the lowest level of concentration, of the test compound which might inhibit the topoisomerase enzyme of interest.
- concentrations Preferably from about three to about eight or more different concentrations of the test compound are tested, the ratio of highest concentration to the lowest concentration being from about 10 ⁇ to about 10 ⁇ .
- concentration of the test compound in the fluid of step (a) is preferably from about 0J ⁇ g/ml to about 1000 ⁇ g/ml, more preferably from about 1 ⁇ g/ml to about 100 ⁇ g/ml.
- the DNA is selected to complement the topoisomerase, in that a DNA that will be changed by interaction with the topoisomerase is needed.
- gyrase a type II topoisomerase
- a relaxed DNA is preferred.
- a suitable, commercially-available relaxed DNA is Relaxed Plasmid pBR 322, available from Lucent Ltd., Leicester, U.K.
- the concentration of the DNA in the fluid of step (a) is preferably from about 0J ⁇ g/ml to about 100 ⁇ g/ml, more preferably from about 1 ⁇ g/ml to about 50 ⁇ g/ml.
- the mole ratio of test compound:DNA in the fluid of step (a) is preferably from about 1 :10" ⁇ to about 1 : 10"3, more preferably from about 1 : 10 ⁇ 6 to about 1 : 10 ⁇ 4.
- the purpose for employing the subject invention methods is generally to identify compounds which will inhibit the topoisomerase of certain organisms of interest, particularly pathogenic bacteria. Compounds which inhibit bacterial topoisomerase may be lethal to such bacteria.
- Type I and type II topoisomerases are known, and are useful in the subject invention methods.
- Gyrase is a preferred type II topoisomerase known to facilitate supercoiling of DNA.
- Commercially available topoisomerases preferred for use in the subject invention methods include the following: "wild-type” gyrase, such as that from E. coli available from Lucent Ltd., and "quinolone-resistance gyrase", such as A(Trp)2B2 available from Lucent Ltd.
- the fluid of step (a) preferably comprises the topoisomerase at a concentration from about 0.001 ⁇ g/ml to about 10 ⁇ g/ml, more preferably from about 0.01 ⁇ g/ml to about 1 ⁇ g/ml.
- the fluid of step (a) preferably has a mole ratio of DNAJopoisomerase of from about 10:1 to about 1 : 100, more preferably from about 1 : 1 to about 1 :10.
- step (b) the fluid from step (a) is incubated to allow the topoisomerase to facilitate changes in the DNA if it is not inhibited from doing so by the test compound.
- the incubation is preferably for a period of from about 0.2 hour to about 24 hours, more preferably from about 1 hour to about 6 hours, preferably at a temperature of from about 20°C to about 55°C, more preferably from about 35°C to about 40°C. 5
- step (c) a cyanine nucleic acid dye is incorporated in the incubated fluid from step (b).
- Preferred cyanine nucleic acid dyes useful in the subject invention assays are disclosed in U.S. Patent Nos. 5,436,134 and 5,658,751 issued to Haugland et al. and Yue et al. on July 25, 1995 and August 19, 1997, respectively, both incorporated herein by reference.
- PicoGreen® is a highly preferred cyanine nucleic acid dye useful in the subject invention methods.
- the dye is added to the incubated fluid from step (b), resulting in a concentration of the dye in the fluid of step (c) of preferably from about 0.01 ⁇ M to about 10 ⁇ M, more preferably from about 0.1 ⁇ M to about 1 ⁇ M.
- the mole ratio of DNA:dye in the fluid of step (c) is preferably from about 1 :10*3 0 a bout 1 :10*7, more preferably from about 1 :10 14 to about 1 :10 16 .
- step (d) the fluorescence from the dye in the fluid from step (c) is measured using known methods.
- the wave lengths or excitation and emission is typically unique for each dye, and is readily ascertained without undue experimentation.
- Preferred for the cyanine dyes useful in the subject invention method is an excitation of from about 470 nm to about 500 nm, more preferably from about 480 nm to about 490 nm, and an emission preferably from about 510 nm to about 540 nm, more preferably from about 520 nm to about 530 nm.
- dithiothreitol 1.8 mMspermidine, 6.5% glycerol (w/v), 0J mg/ml bovine serum albumin), 1.0 ⁇ l of 30 mM ATP, 0.25 ⁇ g of relaxed DNA (Lucent Limited, UK) is placed into each well.
- F(S-DNA) fluorescence measured from wells with supercoiled DNA (to which relaxed DNA and gyrase, but no test compound, are added).
- F(R-DNA) fluorescence measured from wells with relaxed DNA (to which relaxed DNA, but no gyrase or test compound, are added).
- F(TC) fluorescence measured from wells with test compound (to which relaxed DNA, gyrase, and test compound are all added).
- This calculated % Difference should be at least about 30%, or the results from the experiment may not be reliable.
- F(S-DNA) and F(R-DNA) are each typically averages of fluorescence for about four wells.
- F(TC) is typically a single reading of fluorescence for each level of test compound tested.
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP99942591A EP1086242A1 (en) | 1998-03-25 | 1999-03-22 | Fluorescent assay for topoisomerase inhibitors |
JP2000538035A JP2002507433A (en) | 1998-03-25 | 1999-03-22 | Fluorescence assay for topoisomerase inhibitors |
AU32699/99A AU3269999A (en) | 1998-03-25 | 1999-03-22 | Fluorescent assay for topoisomerase inhibitors |
IL13853299A IL138532A0 (en) | 1998-03-25 | 1999-03-22 | Fluorescent assay for topoisomerase inhibitors |
CA002324344A CA2324344A1 (en) | 1998-03-25 | 1999-03-22 | Fluorescent assay for topoisomerase inhibitors |
NO20004753A NO20004753L (en) | 1998-03-25 | 2000-09-22 | Fluorescent Assay for Topoisomerase Inhibitors |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US7926598P | 1998-03-25 | 1998-03-25 | |
US60/079,265 | 1998-03-25 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1999049077A1 true WO1999049077A1 (en) | 1999-09-30 |
Family
ID=22149463
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/IB1999/000473 WO1999049077A1 (en) | 1998-03-25 | 1999-03-22 | Fluorescent assay for topoisomerase inhibitors |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP1086242A1 (en) |
JP (1) | JP2002507433A (en) |
AU (1) | AU3269999A (en) |
CA (1) | CA2324344A1 (en) |
IL (1) | IL138532A0 (en) |
NO (1) | NO20004753L (en) |
WO (1) | WO1999049077A1 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001098540A3 (en) * | 2000-06-22 | 2003-04-24 | Univ State San Diego | Recombination modulators and methods for their production and use |
WO2006051303A1 (en) * | 2004-11-11 | 2006-05-18 | Plant Bioscience Limited | Assay for measuring an enzyme’s capability to modify supercoil topology of nucleic acids and modulators |
WO2010044101A1 (en) * | 2008-10-08 | 2010-04-22 | V. B. Medicare Pvt. Ltd. | Purification and assay for topoisomerases and use of the assay for screening modulators of topoisomerases |
WO2012109617A3 (en) * | 2011-02-10 | 2012-12-06 | University Of Central Florida Research Foundation, Inc. | Method of assaying dna topoisomerases and dna binding proteins using high throughput screening |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996021156A1 (en) * | 1995-01-04 | 1996-07-11 | Abbott Laboratories | Method for preparing scintillation proximity assay targets |
US5759768A (en) * | 1991-05-17 | 1998-06-02 | Dana Farber Cancer Institute | Assays for factors affecting circularization of DNA, assays for factors affecting DNA integration, factors, and uses thereof |
-
1999
- 1999-03-22 CA CA002324344A patent/CA2324344A1/en not_active Abandoned
- 1999-03-22 AU AU32699/99A patent/AU3269999A/en not_active Abandoned
- 1999-03-22 JP JP2000538035A patent/JP2002507433A/en not_active Withdrawn
- 1999-03-22 WO PCT/IB1999/000473 patent/WO1999049077A1/en not_active Application Discontinuation
- 1999-03-22 EP EP99942591A patent/EP1086242A1/en not_active Withdrawn
- 1999-03-22 IL IL13853299A patent/IL138532A0/en unknown
-
2000
- 2000-09-22 NO NO20004753A patent/NO20004753L/en not_active Application Discontinuation
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5759768A (en) * | 1991-05-17 | 1998-06-02 | Dana Farber Cancer Institute | Assays for factors affecting circularization of DNA, assays for factors affecting DNA integration, factors, and uses thereof |
WO1996021156A1 (en) * | 1995-01-04 | 1996-07-11 | Abbott Laboratories | Method for preparing scintillation proximity assay targets |
Non-Patent Citations (1)
Title |
---|
OSBURNE, MARCIA S. ET AL: "An assay for the detection of bacterial DNA gyrase inhibitors", J. ANTIBIOT. (1993), 46(11), 1764-6 CODEN: JANTAJ;ISSN: 0021-8820, XP002107170 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001098540A3 (en) * | 2000-06-22 | 2003-04-24 | Univ State San Diego | Recombination modulators and methods for their production and use |
WO2006051303A1 (en) * | 2004-11-11 | 2006-05-18 | Plant Bioscience Limited | Assay for measuring an enzyme’s capability to modify supercoil topology of nucleic acids and modulators |
US7838230B2 (en) | 2004-11-11 | 2010-11-23 | Plant Bioscience Limited | Assay for measuring an enzyme's capability to modify supercoil topology of nucleic acids and modulators |
WO2010044101A1 (en) * | 2008-10-08 | 2010-04-22 | V. B. Medicare Pvt. Ltd. | Purification and assay for topoisomerases and use of the assay for screening modulators of topoisomerases |
WO2012109617A3 (en) * | 2011-02-10 | 2012-12-06 | University Of Central Florida Research Foundation, Inc. | Method of assaying dna topoisomerases and dna binding proteins using high throughput screening |
Also Published As
Publication number | Publication date |
---|---|
CA2324344A1 (en) | 1999-09-30 |
NO20004753L (en) | 2000-11-23 |
NO20004753D0 (en) | 2000-09-22 |
JP2002507433A (en) | 2002-03-12 |
AU3269999A (en) | 1999-10-18 |
EP1086242A1 (en) | 2001-03-28 |
IL138532A0 (en) | 2001-10-31 |
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