WO2000067558B1 - Regulation of gene expression in tobacco for manipulation of plant growth and secondary metabolism - Google Patents
Regulation of gene expression in tobacco for manipulation of plant growth and secondary metabolismInfo
- Publication number
- WO2000067558B1 WO2000067558B1 PCT/US2000/012450 US0012450W WO0067558B1 WO 2000067558 B1 WO2000067558 B1 WO 2000067558B1 US 0012450 W US0012450 W US 0012450W WO 0067558 B1 WO0067558 B1 WO 0067558B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- vector
- dna molecule
- isolated dna
- sequence
- Prior art date
Links
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract 13
- 241000208125 Nicotiana Species 0.000 title claims 9
- 230000008635 plant growth Effects 0.000 title 1
- 230000014493 regulation of gene expression Effects 0.000 title 1
- 230000024053 secondary metabolic process Effects 0.000 title 1
- 239000002773 nucleotide Substances 0.000 claims abstract 9
- 125000003729 nucleotide group Chemical group 0.000 claims abstract 9
- 230000009261 transgenic effect Effects 0.000 claims abstract 8
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract 7
- 108090000623 proteins and genes Proteins 0.000 claims abstract 7
- 241000196324 Embryophyta Species 0.000 claims abstract 6
- 102000004169 proteins and genes Human genes 0.000 claims abstract 6
- 244000061176 Nicotiana tabacum Species 0.000 claims abstract 4
- 229930013930 alkaloid Natural products 0.000 claims abstract 4
- 230000000295 complement effect Effects 0.000 claims abstract 4
- 150000003797 alkaloid derivatives Chemical class 0.000 claims abstract 3
- 230000015572 biosynthetic process Effects 0.000 claims abstract 3
- 239000012634 fragment Substances 0.000 claims abstract 3
- 230000014509 gene expression Effects 0.000 claims abstract 3
- 108020004414 DNA Proteins 0.000 claims 12
- 102000053602 DNA Human genes 0.000 claims 12
- 108020004999 messenger RNA Proteins 0.000 claims 4
- 108020005544 Antisense RNA Proteins 0.000 claims 3
- 238000000034 method Methods 0.000 claims 3
- 150000007523 nucleic acids Chemical class 0.000 claims 3
- 238000013518 transcription Methods 0.000 claims 3
- 230000035897 transcription Effects 0.000 claims 3
- 102000039446 nucleic acids Human genes 0.000 claims 2
- 108020004707 nucleic acids Proteins 0.000 claims 2
- 108091034117 Oligonucleotide Proteins 0.000 claims 1
- 239000002253 acid Substances 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 239000003184 complementary RNA Substances 0.000 claims 1
- 230000001105 regulatory effect Effects 0.000 claims 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 abstract 2
- 102000007357 Methionine adenosyltransferase Human genes 0.000 abstract 2
- 108010007784 Methionine adenosyltransferase Proteins 0.000 abstract 2
- 102000052812 Ornithine decarboxylases Human genes 0.000 abstract 2
- 108700005126 Ornithine decarboxylases Proteins 0.000 abstract 2
- 230000000692 anti-sense effect Effects 0.000 abstract 2
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 abstract 1
- 102100032252 Antizyme inhibitor 2 Human genes 0.000 abstract 1
- 101100244387 Candida albicans (strain SC5314 / ATCC MYA-2876) PMT6 gene Proteins 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 abstract 1
- 101000798222 Homo sapiens Antizyme inhibitor 2 Proteins 0.000 abstract 1
- 102000006746 NADH Dehydrogenase Human genes 0.000 abstract 1
- 108010086428 NADH Dehydrogenase Proteins 0.000 abstract 1
- 101150036326 PMT2 gene Proteins 0.000 abstract 1
- 101150024216 PMT3 gene Proteins 0.000 abstract 1
- 101150086142 PMT4 gene Proteins 0.000 abstract 1
- 108010086950 Phosphoribosylanthranilate isomerase Proteins 0.000 abstract 1
- 108091022910 Putrescine N-methyltransferase Proteins 0.000 abstract 1
- 101100434411 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) ADH1 gene Proteins 0.000 abstract 1
- 101100241858 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) OAC1 gene Proteins 0.000 abstract 1
- 101100043108 Schizosaccharomyces pombe (strain 972 / ATCC 24843) spb1 gene Proteins 0.000 abstract 1
- 101150102866 adc1 gene Proteins 0.000 abstract 1
- 101150042711 adc2 gene Proteins 0.000 abstract 1
- 238000003556 assay Methods 0.000 abstract 1
- SNICXCGAKADSCV-UHFFFAOYSA-N nicotine Natural products CN1CCCC1C1=CC=CN=C1 SNICXCGAKADSCV-UHFFFAOYSA-N 0.000 abstract 1
- 229960002715 nicotine Drugs 0.000 abstract 1
- 101150092906 pmt1 gene Proteins 0.000 abstract 1
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
- C12N9/0036—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on NADH or NADPH (1.6)
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- C—CHEMISTRY; METALLURGY
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
- C12N15/8223—Vegetative tissue-specific promoters
- C12N15/8227—Root-specific
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
- C12N15/823—Reproductive tissue-specific promoters
- C12N15/8231—Male-specific, e.g. anther, tapetum, pollen
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1003—Transferases (2.) transferring one-carbon groups (2.1)
- C12N9/1007—Methyltransferases (general) (2.1.1.)
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
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- C12Y—ENZYMES
- C12Y401/00—Carbon-carbon lyases (4.1)
- C12Y401/01—Carboxy-lyases (4.1.1)
- C12Y401/01017—Ornithine decarboxylase (4.1.1.17)
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
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- General Engineering & Computer Science (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
This invention relates to enzymes involved in alkaloid, and specifically nicotine, formation in tobacco plants. The invention is based, at least in part, on the nucleotide sequences encoding four variants of putrescine N-methyltransferase (PMT1, PMT2, PMT3, and PMT4), two variants of arginine decarboxylase (ADC1 and ADC2), ornithine decarboxylase (ODC), S-adenosylmethionine synthetase (SAMS), a fragment of NADH dehydrogenase, and a fragment of phosphoribosylanthranilate isomerase. The invention also relates to proteins expressed by these nucleotides, promoter regions of these nucleotides, use of these promoter regions to culture transgenic plant cells and to produce transgenic plants, sense and antisense nucleotides complementary to all or portions of these nucleotide sequences, use of sense and antisense nucleotides to regulate gene expression, and assays using proteins involved in alkaloid formation in tobacco plants.
Claims
AMENDED CLAIMS
[received by the International Bureau on 6 November 2000 (06 11 00), original claim 9 amended, remaining claims unchanged (2 pages)]
1 An isolated DNA molecule comprising the nucleotide sequence of (SEQ. ID. NO. 2), (SEQ ED NO. 5), (SEQ ID NO 8), (SEQ. ID. NO. 11), (SEQ. ID NO 13), (SEQ. ID. NO. 15), (SEQ ID NO IS), (SEQ. ID. NO. 21), (SEQ. ID. NO. 23), (SEQ. ID. NO 25) or (SEQ. ID. NO. 26) or comprising a nucleotide sequence encoding the amino acid sequence encoded by (SEQ ID NO 3), (SEQ. ID NO 6), (SEQ ID. NO 9), (SEQ ID. NO. 12), (SEQ. ID. NO 14), (SEQ. ID. NO 16), (SEQ ID NO. 19), (SEQ. ED NO 22) OR (SEQ. ID. NO. 24)
2. A vector comprising the isolated DNA molecule of claim 1 operably linked to sequences capable of directing the transcπpnon of a mRNA encoded by said isolated DNA molecule
3. An isolated D A molecule comprising a DNA sequence complementary to the nucleotide sequence of claim 1.
4 A vector comprising the isolated DNA molecule of claim 3 operably linked to sequences capable of directing the transcription of a mRNA encoded by said isolated DNA molecule.
5 A cultured iransgenic tobacco cell stably transformed with the vector of claim 2
6. A cultured transgenic tobacco cell stably transformed with the vector of claim 4
7. A transgenic tobacco plant stably transformed with the vector of claim 2.
8. A transgenic iobacco plant stably transformed with the vector of claim 4
9 The isolaied DNA molecule of claim 1, wherein the isolated DNA molecule comprises the nucleotide sequence of (SEQ. ID. NO. 2), (SEQ. ID. NO. 5), (SEQ. ID NO. 8), (SEQ ID NO. 11), (SEQ. ID. NO 13), (SEQ ID. NO. 15), (SEQ. ID. NO 18), (SEQ. ID. NO 21), (SEQ. ID NO 23), (SEQ ID. NO 25) or (SEQ. ID NO. 26)
10 A vector comprising the isolated DNA molecule αf claim 9 operably linked to sequences capable of directing the transcription of a mRNA encoded by said isolated DNA molecule.
11. An isolated DNA molecule comprising a DNA sequence complementary to the nucleotide sequence of the isolated DNA molecule of claim 9.
12. An isolated DNA sequence comprising about a fifteen to about a twenty-five base pair oligonucleotide sequence identical to any consecutive about fifteen to about twenty-five base pair sequence found in (SEQ. ID. NO. 2), (SEQ. ID. NO. 5), (SEQ. ID. NO. 8), (SEQ ID. NO. 11),
(SEQ. ID NO. 13), (SEQ. ID. NO. 15), (SEQ. ID. NO. 18), (SEQ. ID. NO. 21), (SEQ ID NO. 23), (SEQ ID. NO. 25) or (SEQ. ED. NO. 26).
13 A cultured transgenic tobacco cell stably transformed with the vector of claim 10.
14 A transgenic tobacco plant stably transformed with the vector of claim 10.
15 A vector comprising a DNA sequence which encodes an antisense mRNA which is complementary to a fragment of a mRNA encoded by the isolated DNA molecule of claim 1, wherem said sequence is operably linked to sequences capable of directing the transcription of said antisense mRNA in tobacco cells and wherein the expression of said antisense RNA in tobacco cells is sufficient to provide for reduced niconne content m tobacco cells which are stably transformed with said vector as compared to untransformed tobacco cells.
16. A cultured transgenic tobacco cell stably transformed with the vector of claim 15.
17. An isolated and puπfied protein comprising the ammo acid sequence identified m (SEQ ID NO 3), (SEQ. ID. NO. 6), (SEQ ID. NO. 9), (SEQ. ID. NO. 12), (SEQ. ID. NO. 14), (SEQ. ID. NO. 16), (SEQ ID NO 19), (SEQ. ID. NO. 22) or (SEQ. ID. NO. 24).
18. A method for regulating gene expression in a plant comprising funcuonally linking an alkaloid gene promoter to a nucleic acid encoding a protein, wherein the promoter comprises a nucleic acid sequence selected from the group consisnπg of the sequences identified m (SEQ ID NO. 1), (SEQ ID NO. 4), (SEQ ID. NO. 7), (SEQ. ID. NO. 10), (SEQ. ID. NO. I7),and (SEQ. ID. NO. 20)
19. The method of claim 18, wherein the nucleic acid encoding a protein encodes a protein involved in the biosynthesis of alkaloids in plants.
20 A plant transformed by the method of claim 18
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU49915/00A AU4991500A (en) | 1999-05-06 | 2000-05-05 | Regulation of gene expression in tobacco for manipulation of plant growth and secondary metabolism |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13291999P | 1999-05-06 | 1999-05-06 | |
| US60/132,919 | 1999-05-06 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO2000067558A1 WO2000067558A1 (en) | 2000-11-16 |
| WO2000067558B1 true WO2000067558B1 (en) | 2001-02-01 |
Family
ID=22456169
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2000/012450 WO2000067558A1 (en) | 1999-05-06 | 2000-05-05 | Regulation of gene expression in tobacco for manipulation of plant growth and secondary metabolism |
Country Status (2)
| Country | Link |
|---|---|
| AU (1) | AU4991500A (en) |
| WO (1) | WO2000067558A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6907887B2 (en) | 2001-06-08 | 2005-06-21 | Vector Tobacco Ltd. | Modifying nicotine and nitrosamine levels in tobacco |
| US6911541B2 (en) | 2000-08-30 | 2005-06-28 | North Carolina State University | Promoter fragment that is recognized by the product of the tobacco Nic gene |
| US7189570B2 (en) | 2000-11-07 | 2007-03-13 | North Carolina State University | Putrescine-n-methyltransferase promoter |
| US7304220B2 (en) | 1997-06-12 | 2007-12-04 | North Carolina State University | Regulation of quinolate phosphoribosyl transferase expression |
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| WO2002023974A1 (en) | 2000-09-20 | 2002-03-28 | Toyobo Research Center Co., Ltd. | Plant having improved tolerance to various environmental stresses, method of constructing the same and polyamine metabolism-relating enzyme gene |
| CN1954688A (en) * | 2001-06-06 | 2007-05-02 | 二十二世纪有限责任公司 | Tobacco biomass utilization |
| JP3775506B2 (en) | 2002-04-08 | 2006-05-17 | 東洋紡績株式会社 | Plant with improved organ formation and method for producing the same |
| KR20060032141A (en) * | 2003-06-04 | 2006-04-14 | 벡터 토바코 리미티드 | How to reduce the harmful effects of nicotine delivered orally or transdermally |
| US9814258B2 (en) | 2003-08-19 | 2017-11-14 | 22Nd Century Limited, Llc | Reduced-exposure tobacco products |
| WO2006091194A1 (en) | 2005-02-23 | 2006-08-31 | North Carolina State University | Alteration of tobacco alkaloid content through modification of specific cytochrome p450 genes |
| AU2006247739A1 (en) | 2005-05-11 | 2006-11-23 | Vector Tobacco Inc. | Reduced risk tobacco products and methods of making same |
| EP2520655A3 (en) | 2007-07-13 | 2012-12-26 | BASF Plant Science GmbH | Transgenic plants with increased stress tolerance and yield |
| WO2009035546A2 (en) * | 2007-09-07 | 2009-03-19 | University Of Virginia Patent Foundation | Compositions and related methods for modulating transcriptional activation by incorporating gag motifs upstream of core promoter elements |
| BRPI0820042B1 (en) | 2007-11-12 | 2020-05-19 | North Carolina State University | method of obtaining a tobacco plant, or cell or part thereof, having reduced levels of nornicotine, tobacco product, method of making a tobacco product, isolated polynucleotide, expression cassette, isolated polypeptide and method of obtaining a plant, or part of this plant, of the genus nicotiana |
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| CN116250483B (en) | 2015-06-26 | 2024-01-30 | 奥驰亚客户服务公司 | Compositions and methods for producing tobacco plants and articles having altered alkaloid content |
| CN105420258B (en) * | 2015-12-24 | 2019-03-01 | 中国烟草总公司郑州烟草研究院 | The cloning process of tobacco ATP synthase reference gene and its application |
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| WO2023036691A1 (en) * | 2021-09-10 | 2023-03-16 | Philip Morris Products S.A. | Modulating alkaloid profiles in nicotiana tabacum |
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-
2000
- 2000-05-05 WO PCT/US2000/012450 patent/WO2000067558A1/en active Application Filing
- 2000-05-05 AU AU49915/00A patent/AU4991500A/en not_active Abandoned
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7304220B2 (en) | 1997-06-12 | 2007-12-04 | North Carolina State University | Regulation of quinolate phosphoribosyl transferase expression |
| US7408098B2 (en) | 1997-06-12 | 2008-08-05 | North Carolina State University | Regulation of quinolate phosphoribosyl transferase expression |
| US7425670B2 (en) | 1997-06-12 | 2008-09-16 | North Carolina State University | Methods and compositions for protein production in tobacco plants with reduced nicotine |
| US7605308B2 (en) | 1997-06-12 | 2009-10-20 | North Carolina State University | Regulation of quinolate phosphoribosyl transferase expression |
| US6911541B2 (en) | 2000-08-30 | 2005-06-28 | North Carolina State University | Promoter fragment that is recognized by the product of the tobacco Nic gene |
| US7192771B2 (en) | 2000-08-30 | 2007-03-20 | North Carolina State University | Plant promoter sequence |
| US7189570B2 (en) | 2000-11-07 | 2007-03-13 | North Carolina State University | Putrescine-n-methyltransferase promoter |
| US6907887B2 (en) | 2001-06-08 | 2005-06-21 | Vector Tobacco Ltd. | Modifying nicotine and nitrosamine levels in tobacco |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2000067558A1 (en) | 2000-11-16 |
| AU4991500A (en) | 2000-11-21 |
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