- Document Number:
20050119306
- Appl. No:
10/943331
- Application Filed:
September 17, 2004
- نبذة مختصرة :
The present invention provides an alkaloid compound that is an inhibitor of mycotoxin biosynthesis. The alkaloid is an alkenyl piperidine amide wherein the alkenyl is a C18 alkenyl with one or more double bonds. The alkaloid inhibits transcription of the fungus genes nor-1, tri5, and ipnA. The present invention further provides a method for identifying compounds, which inhibit biosynthesis of aflatoxin in Aspergillus spp. and deoxynivalenol in Gibberella spp. without inhibiting growth of the fungus in vitro.
- Inventors:
Trail, Frances (Mason, MI, US); Hammerschmidt, Raymond (East Lansing, MI, US); Linz, John E. (East Lansing, MI, US); Xu, Haixin (East Lansing, MI, US); Velasquez, Luis (Lansing, MI, US); Annis, Seanna L. (Bangor, ME, US)
- Assignees:
Board of Trustees of Michigan State University (East Lansing, MI, US)
- Claim:
1-23. (canceled)
- Claim:
24. A method for determining whether a compound inhibits biosynthesis of a secondary metabolite comprising: (a) providing a culture of a transgenic fungus comprising a reporter gene operably linked to a promoter for a gene involved in the biosynthesis of the secondary metabolite; (b) providing to the culture the compound to be determined; (c) incubating the culture containing the compound under conditions that cause biosynthesis of the secondary metabolite; and (d) measuring expression of the reporter gene wherein absence of the expression of the reporter gene indicates that the compound inhibits biosynthesis of the secondary metabolite.
- Claim:
25. The method of claim 24 wherein the secondary metabolite is a mycotoxin.
- Claim:
26. The method of claim 25 wherein the mycotoxin is selected from the group consisting of aflatoxin, deoxynivalenol, or sterigmatocystin.
- Claim:
27. The method of claim 24 wherein the transgenic fungus is made from a fungus selected from the group consisting of Aspergillus parasiticus, Aspergillus nidulans, Aspergillus versicolor, Aspergillus flavus, Gibberella pulicaris, and Gibberella zeae.
- Claim:
28. The method of claim 24 wherein the promoter is selected from the group consisting of a nor-1 promoter, a ver-1 promoter, verA promoter, fas-1A promoter, omt-1 promoter, an alfR promoter, an ipnA promoter, a tri5 promoter, and mutant thereof.
- Claim:
29. The method of claim 24 wherein the reporter gene is selected from the group consisting of a gene encoding β-glucuronidase, a gene encoding β-galactosidase, a gene encoding luciferase, and a gene encoding fluorescence green protein.
- Claim:
30. The method of claim 24 wherein a transgenic fungus is provided that contains a reporter gene operatively linked to a constitutive promoter which provides a control for the method.
- Claim:
31. The method of claim 30 wherein the constitutive promoter is benA or mutant thereof.
- Claim:
32. A method for identifying a compound in a material that inhibits the biosynthesis of a secondary metabolite of a fungus comprising: (a) providing an extract of the material; (b) separating the material by a chromatography method; (c) providing a spore suspension of a transgenic fungus comprises a reporter gene operatively linked to a promoter that is the same as the promoter that controls transcription of a gene involved in biosynthesis of the secondary metabolite; (d) adding the spore suspension to the separated material; (e) allowing the spores to germinate and grow fungi; and (f) detecting expression of the reporter gene wherein absence of the expression of the reporter gene identifies the separated material that inhibits the biosynthesis of the secondary metabolite.
- Claim:
33. The method of claim 32 wherein the secondary metabolite is a mycotoxin.
- Claim:
34. The method of claim 33 wherein the mycotoxin is selected from the group consisting of aflatoxin, deoxynivalenol, and sterigmatocystin.
- Claim:
35. The method of claim 32 wherein the chromatography is thin layer chromatography (TLC) using TLC plates.
- Claim:
36. The method of claim 32 wherein the transgenic fungus is a fungus selected from the group consisting of Aspergillus parasiticus, Aspergillus nidulans, Aspergillus versicolor, Aspergillus flavus, Gibberella pulicaris and Gibberella zeae.
- Claim:
37. The method of claim 32 wherein the promoter is selected from the group consisting of a nor-1 promoter, a ver-1 promoter, verA promoter, fas-1a promoter, omt-1 promoter, an alfR promoter, an ipnA promoter, a tri5 promoter, and mutant thereof.
- Claim:
38. The method of claim 32 wherein the reporter gene is selected from the group consisting of a gene encoding β-glucuronidase, a gene encoding β-galactosidase, a gene encoding luciferase, and a gene encoding fluorescent green protein.
- Claim:
39. The method of claim 32 wherein a transgenic fungus is provided that contains a reporter gene operatively linked to a constitutive promoter which provides a control for the method.
- Claim:
40. The method of claim 39 wherein the constitutive promoter is benA or mutant thereof.
- Claim:
41. The method of claim 35 wherein the spores are applied to the TLC plates in an agarose solution.
- Claim:
42. The method of claim 32 or 35 wherein the fungi are incubated in a dark and moist atmosphere at 30° C. for a time sufficient for the fungi to cover the plate.
- Claim:
43. The method of claim 32 or 35 wherein the fungi are lysed to release the reporter gene expression product.
- Claim:
44. The method of claim 43 wherein the fungi are lysed by freeze-thawing.
- Current U.S. Class:
514317/000
- الرقم المعرف:
edspap.20050119306
Processing Request
No Comments.