Novel protein, genes encoding the same and method of using the same
AssigneeJapan Tobacco Inc.
InventorYoshimitsu Takakura
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Abstract
An object of the present invention is to search and identify novel antifungal proteins capable of inhibiting the growth of plant pathogenic microorganisms including Magnaporthe grisea and Rhizoctonia solani causing two major rice diseases at relatively low concentrations, and further to clone a gene for said protein. The present invention provides an antifungal protein which can be obtained from fraction(s) precipitated by ammonium sulfate precipitation using an aqueous extract from Pleurotus cornucopiae, wherein said protein has an antifungal activity against at least rice blast, and exhibits existence of a component having a molecular weight of about 15 kDa as determined by SDS-PAGE method; a gene encoding said protein and uses thereof.
Claims26
An antifungal protein which can be obtained from fraction(s) precipitated by ammonium sulfate precipitation method using aqueous extract from Pleurotus cornucopiae, wherein said protein has an antifungal activity against at least rice blast, and exhibits existence of a component having a molecular weight of about 15 kDa by SDS-PAGE method.
An antifungal protein exhibiting an antifungal activity against rice blast, wherein said protein has an amino acid sequence of the amino acid sequence of SEQ ID NO:2 or NO:4, or an amino acid sequence having one or more amino acid modifications in said sequences, or an amino acid with 52% or more homology to said sequences.
An antifungal protein consisting of a polypeptide alone or a combination thereof, wherein said polypeptide is selected from
- a polypeptide consisting of the partial amino acid sequence 8-143 of SEQ ID NO:2 or the partial amino acid sequence 8-141 of SEQ ID NO:4, and
- a polypeptide having one or more amino acid modifications in one of said amino acid sequences, and
- a polypeptide having an amino acid sequence with 52% or more homology to any one of said amino acid sequences
- wherein said polypeptides exhibit an antifungal activity against rice blast.
An oligonucleotide for obtaining a gene encoding an antifungal protein from Pleurotus cornucopiae, produced by a method, which comprises:
- selecting two regions from a base sequence of a gene encoding the antifungal protein of SEQ ID NO:l or NO:3 based on the following requirements;
- 1) length of each regions is 15-30 bases;
- 2) proportion of G+C content in a base sequence of each region is 40-60%;
- preparing a single-stranded DNA having a base sequence which is identical to said region or complementary to said region, or preparing mixture of single-stranded DNAs based on degeneracy of the genetic code without changing a sequence of amino acid residues encoded by said single-stranded DNAs;
- and
- optionally preparing a modified version of said single-stranded DNAs, said modification not altering a binding specificity of the single-stranded DNAs to the base sequence of the gene encoding said antifungal protein.
Dependencies are inferred from the claim text.