Detection of a Point Mutation in FAS2 Gene of Sake Yeast Strains by Allele-Specific PCR Amplification

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概要

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• To identify yeast mutants with a point mutation, detection of the specific mutant alleles is necessary. For this purpose, we applied allele-specific polymerase chain reaction (PCR) to detect the FAS2-1250S dominant mutant allele that encodes an altered fatty acid synthase in Japanese brewer's yeast strains. These strains are known to produce a higher amount of ethyl caproate in Japanese sake. The mutant strains were supposed to be diploid and to contain heterozygous alleles, including wild-type FAS2 and a dominant FAS2-1250S. A set of oligonucleotide primers was designed to contain different nucleotides at their 3' termini: one type was identical to the wild type and the other to the mutant FAS2. Another set of primers was designed to have an additional mismatch at the second nucleotide from their 3' termini. By testing with control strains, we established PCR conditions for specific amplification. Using these conditions and a simple template preparation procedure with SDS, the presence of the allele was detected in commercially used sake yeast strains. The method presented here will be useful for the identification of specific yeast strains.

• 社団法人日本生物工学会の論文
• 2001-08-25

著者

• HOSHIDA Hisashi

  Department of Applied Molecular Bioscience, Yamaguchi University Graduate School of Medicine

• AKADA Rinji

  Department of Applied Molecular Bioscience, Yamaguchi University Graduate School of Medicine

• Akada Rinji

  Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University

• NISHIZAWA YOSHINORI

  Department of Fermentation Tachnology, Faculty of Engineering, Hiroshima University

• Nishizawa Y

  Department Of Fermentation Technology Faculty Of Engineering Hiroshima University

• Nishizawa Y

  Yamaguchi Univ. Ube Jpn

• Nishizawa Yoshinori

  Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University

• Nishizawa Yoshinori

  Department Of Fermentation Technology Faculty Of Engineering Hiroshima University

• Hoshida Hisashi

  Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University

• HIROSAWA Isao

  Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi Universi

• Hirosawa Isao

  Department Of Applied Chemistry And Chemical Engineering Faculty Of Engineering Yamaguchi University

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