Transformed cell with enhanced sensitivity to antifungal compound and use thereof
AssigneeSumitomo Chemical Company, Limited
InventorHiroki NAKAJIMA
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Abstract
The present invention provides a transformed cell in which a polynucleotide having a nucleotide sequence encoding an amino acid sequence of an osmosensing histidine kinase having no transmembrane region is introduced in a functional form into a cell deficient in at least one hybrid-sensor kinase, a method of assaying the antifungal activity of a test substance using the transformed cell, and a method of searching an antifungal compound using the method, and the like.
Claims19
A transformed cell in which a polynucleotide encoding an osmosensing histidine kinase having no transmembrane region is introduced in a functional form into a cell that is deficient in at least one hybrid-sensor kinase, wherein the osmosensing histidine kinase having no transmembrane region has been obtained from Fusarium oxysporum, Mycospharella tritici or Thanatephorus cucumeris.
A polynucleotide encoding an amino acid sequence selected from the group consisting of:
- (a) an amino acid sequence of an osmosensing histidine kinase having no transmembrane region, which has amino acid sequence homology of 95% or more to the amino acid sequence of any of SEQ ID NOs:
- 41, 55 and 68;
- (b) an amino acid sequence of an osmosensing histidine kinase having no transmembrane region, which is encoded by a DNA amplified by a polymerase chain reaction using a cDNA obtained from Fusarium oxysporum as a template and using as primers an oligonucleotide comprising the nucleotide sequence of SEQ ID NO:
- 52 and an oligonucleotide comprising the nucleotide sequence of SEQ ID NO:
- 53;
- (c) an amino acid sequence of an osmosensing histidine kinase having no transmembrane region, which is encoded by a DNA amplified by a polymerase chain reaction using a cDNA obtained from Mycospharella tritici as a template and using as primers an oligonucleotide comprising the nucleotide sequence of SEQ ID NO:
- 64 and an oligonucleotide comprising the nucleotide sequence of SEQ ID NO:
- 65;
- (d) an amino acid sequence of an osmosensing histidine kinase having no transmembrane region, which is encoded by a DNA amplified by a polymerase chain reaction using a cDNA obtained from Thanatephorus cucumeris as a template and using as primers an oligonucleotide comprising the nucleotide sequence of SEQ ID NO:
- 85 and an oligonucleotide comprising the nucleotide sequence of SEQ ID NO:
- 86;
- (e) the amino acid sequence of SEQ ID NO:
- 41;
- (f) the amino acid sequence of SEQ ID NO:
- 55;
- and
- (g) the amino acid sequence of SEQ ID NO:
- 68.
A method of obtaining from a plant-pathogenic filamentous fungus a polynucleotide encoding an osmosensing histidine kinase having no transmembrane region, which comprises:
- amplifying a desired polynucleotide by polymerase chain reaction using as a primer an oligonucleotide comprising the nucleotide sequence of any of SEQ ID NOs:
- 52, 53, 64, 65, 85 and 86, and recovering the amplified desired polynucleotide.
Dependencies are inferred from the claim text.