Claims
- 1. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) contacting a Chitin Synthase 2 polypeptide with a test compound; and b) detecting the presence or absence of binding between said test compound and said Chitin Synthase 2 polypeptide, wherein binding indicates that said test compound is a candidate for an antibiotic.
- 2. The method of claim 1, wherein said Chitin Synthase 2 polypeptide is a fungal Chitin Synthase 2 polypeptide.
- 3. The method of claim 1, wherein said Chitin Synthase 2 polypeptide is a Magnaporthe Chitin Synthase 2 polypeptide.
- 4. The method of claim 1, wherein said Chitin Synthase 2 polypeptide is SEQ ID NO: 3.
- 5. A method for determining whether the antibiotic candidate of claim 1 has antifungal activity, further comprising: contacting a fungus or fungal cells with said antibiotic candidate and detecting the decrease in growth, viability, or pathogenicity of said fungus or fungal cells.
- 6. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) contacting a test compound with at least one polypeptide selected from the group consisting of: a polypeptide having at least ten consecutive amino acids of a fungal Chitin Synthase 2; a polypeptide having at least 50% sequence identity with a fungal Chitin Synthase 2; and a polypeptide having at least 10% of the activity thereof; and b) detecting the presence and/or absence of binding between said test compound and said polypeptide, wherein binding indicates that said test compound is a candidate for an antibiotic.
- 7. A method for determining whether the antibiotic candidate of claim 6 has antifungal activity, further comprising: contacting a fungus or fungal cells with said antibiotic candidate and detecting a decrease in growth, viability, or pathogenicity of said fungus or fungal cells.
- 8. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) contacting UDP-N-acetyl-D-glucosamine and starter chitin with a Chitin Synthase 2; b) contacting UDP-N-acetyl-D-glucosamine and starter chitin with Chitin Synthase 2 and a test compound; and c) determining the change in concentration for at least one of the following: UDP-N-acetyl-D-glucosamine, starter chitin, extended chitin, and/or UDP, wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.
- 9. The method of claim 8, wherein said Chitin Synthase 2 is a fungal Chitin Synthase 2.
- 10. The method of claim 8, wherein said Chitin Synthase 2 is a Magnaporthe Chitin Synthase 2.
- 11. The method of claim 8, wherein said Chitin Synthase 2 is SEQ ID NO: 3.
- 12. A method for determining whether the antibiotic candidate of claim 8 has antifungal activity, further comprising: contacting a fungus or fungal cells with said antibiotic candidate and detecting a decrease in growth, viability, or pathogenicity of said fungus or fungal cells.
- 13. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) contacting starter chitin and UDP with a Chitin Synthase 2; b) contacting starter chitin and UDP with a Chitin Synthase 2 and a test compound; and c) determining the change in concentration for at least one of the following: UDP-N-acetyl-D-glucosamine, starter chitin, shortened chitin, and/or UDP, wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.
- 14. The method of claim 13, wherein said Chitin Synthase 2 is a fungal Chitin Synthase 2.
- 15. The method of claim 13, wherein said Chitin Synthase 2 is a Magnaporthe Chitin Synthase 2.
- 16. The method of claim 13, wherein said Chitin Synthase 2 is SEQ ID NO: 3.
- 17. A method for determining whether the antibiotic candidate of claim 13 has antifungal activity, further comprising: contacting a fungus or fungal cells with said antibiotic candidate and detecting a decrease in growth, viability, or pathogenicity of said fungus or fungal cells.
- 18. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) contacting UDP-N-acetyl-D-glucosamine and starter chitin with a polypeptide selected from the group consisting of: a polypeptide having at least 50% sequence identity with Chitin Synthase 2; a polypeptide having at least 50% sequence identity with a Chitin Synthase 2 and having at least 10% of the activity thereof; and a polypeptide comprising at least 100 consecutive amino acids of a Chitin Synthase 2; b) contacting UDP-N-acetyl-D-glucosamine and starter chitin with said polypeptide and a test compound; and c) determining the change in concentration for at least one of the following: UDP-N-acetyl-D-glucosamine, starter chitin, extended chitin, and/or UDP, wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.
- 19. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) contacting starter chitin and UDP with a polypeptide selected from the group consisting of: a polypeptide having at least 50% sequence identity with a Chitin Synthase 2; a polypeptide having at least 50% sequence identity with a Chitin Synthase 2 and at least 10% of the activity thereof; and a polypeptide comprising at least 100 consecutive amino acids of a Chitin Synthase 2; b) contacting starter chitin and UDP, with said polypeptide and a test compound; and c) determining the change in concentration for at least one of the following: UDP-N-acetyl-D-glucosamine, starter chitin, shortened chitin, and/or UDP, wherein a change in concentration for any of the above substances between steps (a) and (b) indicates that said test compound is a candidate for an antibiotic.
- 20. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) measuring the expression of a Chitin Synthase 2 in a cell, cells, tissue, or an organism in the absence of a test compound; b) contacting said cell, cells, tissue, or organism with said test compound and measuring the expression of said Chitin Synthase 2 in said cell, cells, tissue, or organism; and c) comparing the expression of Chitin Synthase 2 in steps (a) and (b), wherein a lower expression in the presence of said test compound indicates that said test compound is a candidate for an antibiotic.
- 21. The method of claim 20, wherein said cell, cells, tissue, or organism is, or is derived from a fungus.
- 22. The method of claim 20, wherein said cell, cells, tissue, or organism is, or is derived from a Magnaporthe fungus or fungal cell.
- 23. The method of claim 20, wherein said Chitin Synthase 2 is SEQ ID NO: 3.
- 24. The method of claim 20, wherein the expression of Chitin Synthase 2 is measured by detecting CHS2 mRNA.
- 25. The method of claim 20, wherein the expression of Chitin Synthase 2 is measured by detecting Chitin Synthase 2 polypeptide.
- 26. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) providing cells having one form of a Chitin Synthase 2 gene, and providing comparison cells having a different form of a Chitin Synthase 2 gene; and b) contacting said cells and said comparison cells with a test compound and determining the growth of said cells and comparison cells in the presence of the test compound, wherein a difference in growth between said cells and said comparison cells in the presence of said compound indicates that said compound is a candidate for an antibiotic.
- 27. The method of claim 26, wherein the cells and the comparison cells are fungal cells.
- 28. The method of claim 26, wherein the cells and the comparison cells are Magnaporthe cells.
- 29. The method of claim 26, wherein said form and said different form of the Chitin Synthase 2 are fungal Chitin Synthase 2s.
- 30. The method of claim 26, wherein at least one of the forms is a Magnaporthe Chitin Synthase 2.
- 31. The method of claim 26, wherein said form and said different form of the Chitin Synthase 2 are non-fungal Chitin Synthase 2s.
- 32. The method of claim 26, wherein one form of the Chitin Synthase 2 is a fungal Chitin Synthase 2, and the different form is a non-fungal Chitin Synthase 2.
- 33. A method for identifying a test compound as a candidate for an antibiotic, comprising:
a) providing cells having one form of a gene in the chitin biochemical and/or genetic pathway and providing comparison cells having a different form of said gene. b) contacting said cells and said comparison cells with a said test compound, c) determining the growth of said cells and said comparison cells in the presence of said test compound, wherein a difference in growth between said cells and said comparison cells in the presence of said test compound indicates that said test compound is a candidate for an antibiotic.
- 34. The method of claim 33, wherein the cells and the comparison cells are fungal cells.
- 35. The method of claim 33, wherein the cells and the comparison cells are Magnaporthe cells.
- 36. The method of claim 33, wherein said form and said different form of the chitin biosynthesis gene are fungal chitin biosynthesis genes.
- 37. The method of claim 33, wherein at least one of the forms is a Magnaporthe chitin biosynthesis gene.
- 38. The method of claim 33, wherein said form and said different form of the chitin biosynthesis genes are non-fungal chitin biosynthesis genes.
- 39. The method of claim 33, wherein one form of the chitin biosynthesis gene is a fungal chitin biosynthesis gene, and the different form is a non-fungal chitin biosynthesis gene.
- 40. A method for determining whether the antibiotic candidate of claim 33 has antifungal activity, further comprising: contacting a fungus or fungal cells with said antibiotic candidate and detecting a decrease in growth, viability, or pathogenicity of said fungus or fungal cells, wherein a decrease in growth, viability, or pathogenicity of said fungus or fungal cells indicates that the antibiotic candidate has antifungal activity.
- 41. An isolated nucleic acid comprising a nucleotide sequence that encodes a polypeptide of SEQ ID NO: 3.
- 42. The nucleic acid of claim 41 comprising the nucleotide sequence of SEQ ID NO: 1.
- 43. An expression cassette comprising the nucleic acid of claim 42.
- 44. The isolated nucleic acid of claim 41 comprising a nucleotide sequence with at least 50 to at least 95% sequence identity to SEQ ID NO: 1.
- 45. An isolated polypeptide consisting essentially of the amino acid sequence of SEQ ID NO: 3.
- 46. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 3.
RELATED APPLICATIONS
[0001] This applications claims the benefit of U.S. Application No. 60/381,159 filed May 16, 2002, herein incorporated in its entirety by reference.
Provisional Applications (1)
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Number |
Date |
Country |
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60381159 |
May 2002 |
US |