WO2004050880B1 - Insecticidal agent - Google Patents
Insecticidal agentInfo
- Publication number
- WO2004050880B1 WO2004050880B1 PCT/GB2003/005183 GB0305183W WO2004050880B1 WO 2004050880 B1 WO2004050880 B1 WO 2004050880B1 GB 0305183 W GB0305183 W GB 0305183W WO 2004050880 B1 WO2004050880 B1 WO 2004050880B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- plant according
- protoxin
- modified
- amino acids
- plant
- Prior art date
Links
- 239000002917 insecticide Substances 0.000 title claims 3
- 241000193388 Bacillus thuringiensis Species 0.000 claims abstract 4
- 229940097012 bacillus thuringiensis Drugs 0.000 claims abstract 4
- 230000000749 insecticidal effect Effects 0.000 claims abstract 3
- 241000196324 Embryophyta Species 0.000 claims 19
- 108090000765 processed proteins & peptides Proteins 0.000 claims 5
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 claims 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 150000001413 amino acids Chemical class 0.000 claims 4
- 229920001184 polypeptide Polymers 0.000 claims 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims 4
- 241000500437 Plutella xylostella Species 0.000 claims 3
- 239000000463 material Substances 0.000 claims 3
- 230000017854 proteolysis Effects 0.000 claims 3
- 230000001850 reproductive effect Effects 0.000 claims 3
- 238000006467 substitution reaction Methods 0.000 claims 3
- 241000500439 Plutella Species 0.000 claims 2
- 230000004048 modification Effects 0.000 claims 2
- 238000012986 modification Methods 0.000 claims 2
- 241000894007 species Species 0.000 claims 2
- 241000219193 Brassicaceae Species 0.000 claims 1
- 102000012479 Serine Proteases Human genes 0.000 claims 1
- 108010022999 Serine Proteases Proteins 0.000 claims 1
- 238000003776 cleavage reaction Methods 0.000 claims 1
- 102000038379 digestive enzymes Human genes 0.000 claims 1
- 108091007734 digestive enzymes Proteins 0.000 claims 1
- 230000037406 food intake Effects 0.000 claims 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims 1
- 230000001418 larval effect Effects 0.000 claims 1
- 239000012528 membrane Substances 0.000 claims 1
- 230000006337 proteolytic cleavage Effects 0.000 claims 1
- 230000007017 scission Effects 0.000 claims 1
- 239000003053 toxin Substances 0.000 abstract 2
- 231100000765 toxin Toxicity 0.000 abstract 2
- 108700012359 toxins Proteins 0.000 abstract 2
- 108700003918 Bacillus Thuringiensis insecticidal crystal Proteins 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 abstract 1
- 108091005804 Peptidases Proteins 0.000 abstract 1
- 239000004365 Protease Substances 0.000 abstract 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract 1
- 210000004897 n-terminal region Anatomy 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8286—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for insect resistance
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/32—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Bacillus (G)
- C07K14/325—Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Pest Control & Pesticides (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Insects & Arthropods (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Recombinant Bacillus thuringiensis Cry toxins modified such that the N-terminal region of the protoxin is substantially resistant to protease attack show increased insecticidal activity in lepidopterans resistant to the wild type toxin. The recombinant toxin can be expressed in planta or in bacteria, for instance.
Claims
1. A plant, or reproductive material thereof, transformed with and capable of expressing DNA encoding a Bacillus thuringiensis Cry protoxin, the amino acid sequence of the N-terminus of the protoxin being modified so as to inhibit proteolysis at the said N-terminus.
2. A plant according to claim 1, wherein the expressed Cry protoxin is insecticidal in Plutella sp, after ingestion
3. A plant according to claim 2, wherein the Cry protoxin is insecticidal in Plutella xylostella.
4. A plant according to any preceding claim, wherein the modified protoxin is resistant to serine protease cleavage.
5. A plant according to any preceding claim, wherein the N-terminal of the modified protoxin is substantially stable in the presence of larval Plutella xylostella mid-gut juices.
6. A plant according to any preceding claim, wherein the modified protoxin is susceptible to proteolytic cleavage by the brash border membrane, or vesicles thereof, of Plutella xylostella.
7. A plant according to any preceding claim, wherein at least one position selected from positions 13, 16, 24, 28 and 33, as mapped in SEQ. ID NO: 2, is modified to inhibit proteolysis.
8. A plant according to claim 7, wherein at least residues R28, L24 and Y13 are modified.
9. A plant according to claim 7, wherein all of the said residues are modified.
AMENDED SHEET (ARTICLE 18) 40
10. A plant according to any preceding claim, wherein the protoxin corresponds substantially to Cryl Ac after position 35.
11. A plant according to any preceding claim, wherein the protoxin corresponds substantially to Cryl Ab after position 35.
12. A plant according to any preceding claim, which is a food crop.
13. A plant according to any preceding claim, which is a companion species.
14. A plant according to claim 13, wherein the companion species is of the species Brassicaceae.
15. A plant according to any preceding claim, wherein the modification includes at least one substitution.
16. A plant according to any preceding claim, wherein N terminal amino acids 1- 34 of the Cry protoxin encoded by the DNA have at least 70% amino acid sequence identity with amino acids 1- 34 of SEQ LD NO. 1.
17. A plant according to claim 16, wherein N terminal amino acids 1-34 of the Cry protoxin encoded by the DNA have at least 95 % amino acid sequence identity with amino acids 1- 34 of SEQ ID NO. 2.
18. A plant, or reproductive material thereof, transformed with and capable of expressing DNA encoding the peptide sequence of SEQ ID NO 2.
19. A plant, or reproductive material thereof, transformed with and capable of expressing DNA encoding Cryl Ab but which has the N-terminal amino acid sequence from positions 1 to 35 of SEQ LD NO 2.
20. The use of a modified protoxin as defined in any preceding claim as an insecticide.
AMENDED SHEET (ARTICLE 18) 41
21. A vector comprising DNA as defined in any of claims 1 to 19.
22. A vector according to claim 21, further comprising at least one expression control sequence for the DNA.
23. A host containing a vector according to claim 21 or 22.
24. A host according to claim 23, which is Bacillus thuringiensis.
25. A recombinant Bacillus thuringiensis Cryl Ac polypeptide, for use as an insecticidal agent against Plutella sp., characterised in that the polypeptide comprises a number of substitutions in its N terminal 34 amino acids, wherein the substitutions occur at putative proteolysis sites for digestive enzymes, and wherein N terminal amino acids 1-34 of the recombinant Cryl Ac polypeptide have at least 70% amino acid sequence identity with amino acids 1-34 of SEQ LD NO. 1.
26. A polypeptide according to claim 25, wherein the number of sequence modifications is from 2 to 5.
AMENDED SHEET (ARTICLE 18)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2003285548A AU2003285548A1 (en) | 2002-11-29 | 2003-11-28 | Insecticidal agent |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB0227919.8A GB0227919D0 (en) | 2002-11-29 | 2002-11-29 | Insecticidal agent |
| GB0227919.8 | 2002-11-29 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO2004050880A2 WO2004050880A2 (en) | 2004-06-17 |
| WO2004050880A3 WO2004050880A3 (en) | 2004-07-29 |
| WO2004050880B1 true WO2004050880B1 (en) | 2004-09-16 |
Family
ID=9948806
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/GB2003/005183 WO2004050880A2 (en) | 2002-11-29 | 2003-11-28 | Insecticidal agent |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU2003285548A1 (en) |
| GB (1) | GB0227919D0 (en) |
| WO (1) | WO2004050880A2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BR112017013514A2 (en) * | 2014-12-30 | 2018-01-02 | Dow Agrosciences Llc | modified cry1ca toxins useful for insect pest control |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU6702300A (en) * | 1999-08-19 | 2001-03-19 | Syngenta Participations Ag | Hybrid insecticidal toxins and nucleic acid sequences coding therefor |
-
2002
- 2002-11-29 GB GBGB0227919.8A patent/GB0227919D0/en not_active Ceased
-
2003
- 2003-11-28 WO PCT/GB2003/005183 patent/WO2004050880A2/en not_active Application Discontinuation
- 2003-11-28 AU AU2003285548A patent/AU2003285548A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| WO2004050880A2 (en) | 2004-06-17 |
| AU2003285548A1 (en) | 2004-06-23 |
| AU2003285548A8 (en) | 2004-06-23 |
| GB0227919D0 (en) | 2003-01-08 |
| WO2004050880A3 (en) | 2004-07-29 |
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