WO2003072706A2 - Molecules d'acides nucleiques codant des proteines essentielles a la croissance et au developpement fongiques, et leurs utilisations - Google Patents

Molecules d'acides nucleiques codant des proteines essentielles a la croissance et au developpement fongiques, et leurs utilisations Download PDF

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Publication number
WO2003072706A2
WO2003072706A2 PCT/US2003/004154 US0304154W WO03072706A2 WO 2003072706 A2 WO2003072706 A2 WO 2003072706A2 US 0304154 W US0304154 W US 0304154W WO 03072706 A2 WO03072706 A2 WO 03072706A2
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WO
WIPO (PCT)
Prior art keywords
seq
acid sequence
amino acid
protein
nucleic acid
Prior art date
Application number
PCT/US2003/004154
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English (en)
Other versions
WO2003072706A3 (fr
Inventor
Thomas Deane Gaffney
Albert Flavier
Frederick Dietrich
Peter Philippsen
Cloyd M. Michelle Kirksey
Krista Gates
Joann Gardner
Original Assignee
Syngenta Participations Ag
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Syngenta Participations Ag filed Critical Syngenta Participations Ag
Priority to AU2003225561A priority Critical patent/AU2003225561A1/en
Publication of WO2003072706A2 publication Critical patent/WO2003072706A2/fr
Publication of WO2003072706A3 publication Critical patent/WO2003072706A3/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi

Definitions

  • the critical role in fungal growth of the proteins encoded by each of the nucleotide sequences of the invention implies that chemicals that inhibit the function of any one of these proteins in fungi are likely to have detrimental effects on fungi and are potentially good fungicide candidates.
  • the proteins encoded by the essential nucleotide sequences provide the bases for assays designed to easily and rapidly identify novel fungicides.
  • the present invention provides a method of identifying a fungicidal compound, comprising: (a) combining a polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of the even numbered SEQ ID NOs:2-60 with a compound to be tested for the ability to bind to said polypeptide, under conditions conducive to binding; (b) selecting a compound identified in (a) that binds to said polypeptide; (c) applying a compound selected in (b) to a plant to test for fungicidal activity; and (d) selecting a compound identified in (c) that has fungicidal activity.
  • the polypeptide comprises an amino acid sequence at least 95 % identical to an amino acid sequence selected from the group consisting of the even numbered SEQ ID NOs:2-60.
  • SEQ ID NOs:61-124 are oligonucleotide primers. Table 1 Fungal Essential Genes and Identification
  • Complementary refers to two nucleotide sequences that comprise antiparallel nucleotide sequences capable of pairing with one another upon formation of hydrogen bonds between the complementary base residues in the antiparallel nucleotide sequences.
  • Enzyme activity means herein the ability of an enzyme to catalyze the conversion of a substrate into a product.
  • a substrate for the enzyme comprises the natural substrate of the enzyme but also comprises analogues of the natural substrate, which can also be converted, by the enzyme into a product or into an analogue of a product. The activity of the enzyme is measured for example by determining the amount of product in the reaction after a certain period of time, or by determining the amount of substrate remaining in the reaction mixture after a certain period of time.
  • Percent identity means open reading frame.
  • Percent identity the phrases “percent identical” or “percent identical,” in the context of two nucleic acid or protein sequences, refers to two or more sequences or subsequences that have for example 60%, preferably 70%, more preferably 80%, still more preferably 90%, even more preferably 95%, and most preferably at least 99% nucleotide or amino acid residue identity, when compared and aligned for maximum correspondence, as measured using one of the following sequence comparison algorithms or by visual inspection.
  • the percent identity exists over a region of the sequences that is at least about 50 residues in length, more preferably over a region of at least about 100 residues, and most preferably the percent identity exists over at least about 150 residues. In an especially prefened embodiment, the percent identity exists over the entire length of the coding regions.
  • nucleic acid or protein is at least about 50% pure, more preferably at least about 85% pure, and most preferably at least about 99% pure.
  • Two nucleic acids are "recombined” when sequences from each of the two nucleic acids are combined in a progeny nucleic acid.
  • Two sequences are “directly” recombined when both of the nucleic acids are substrates for recombination.
  • Two sequences are "indirectly recombined” when the sequences are recombined using an intermediate such as a cross-over oligonucleotide. For indirect recombination, no more than one of the sequences is an actual substrate for recombination, and in some cases, neither sequence is a substrate for recombination.
  • a simple assay is developed to screen for chemicals that affect normal functioning of the polypeptides of the invention. Such chemicals are promising in vitro leads that can be tested for in vivo fungicidal activity.
  • Any one of the nucleotide sequences of the invention is operatively linked to a strong inducible promoter, such promoters being known in the art.
  • the vector comprising the selected nucleotide of the invention operatively linked to the selected inducible promoter is transformed into a host, such as E. coli. Transformed E.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Mycology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

L'invention concerne des séquences de nucléotides isolées de Ashbya gossypii qui codent des protéines essentielles à la croissance et au développement fongiques. On peut utiliser l'essentiel des protéines en exprimant par recombinaison les protéines et en les utilisant dans des dosages de criblage, de manière à identifier des composés qui interagissent ou inhibent les protéines et, de ce fait, sont fongicides.
PCT/US2003/004154 2002-02-22 2003-02-11 Molecules d'acides nucleiques codant des proteines essentielles a la croissance et au developpement fongiques, et leurs utilisations WO2003072706A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003225561A AU2003225561A1 (en) 2002-02-22 2003-02-11 Nucleic acid molecules encoding proteins essential for fungal growth and development and uses thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US35955702P 2002-02-22 2002-02-22
US60/359,557 2002-02-22

Publications (2)

Publication Number Publication Date
WO2003072706A2 true WO2003072706A2 (fr) 2003-09-04
WO2003072706A3 WO2003072706A3 (fr) 2003-12-24

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PCT/US2003/004154 WO2003072706A2 (fr) 2002-02-22 2003-02-11 Molecules d'acides nucleiques codant des proteines essentielles a la croissance et au developpement fongiques, et leurs utilisations

Country Status (2)

Country Link
AU (1) AU2003225561A1 (fr)
WO (1) WO2003072706A2 (fr)

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
GOFFEAU ET AL.: 'Life with 6000 genes' SCIENCE MAGAZINE vol. 274, 25 October 1996, pages 546 - 567, XP002970778 *

Also Published As

Publication number Publication date
WO2003072706A3 (fr) 2003-12-24
AU2003225561A8 (en) 2003-09-09
AU2003225561A1 (en) 2003-09-09

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