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Titolo:
Detection of a point mutation in FAS2 gene of sake yeast strains by allele-specific PCR amplification
Autore:
Akada, R; Hirosawa, I; Hoshida, H; Nishizawa, Y;
Indirizzi:
Yamaguchi Univ, Fac Engn, Dept Chem Engn & Appl Chem, Ube, Yamaguchi 7558611, Japan Yamaguchi Univ Ube Yamaguchi Japan 7558611 Ube, Yamaguchi 7558611, Japan
Titolo Testata:
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
fascicolo: 2, volume: 92, anno: 2001,
pagine: 189 - 192
SICI:
1389-1723(200108)92:2<189:DOAPMI>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
SICKLE-CELL-ANEMIA; SACCHAROMYCES-CEREVISIAE; DNA; MUTANTS; ACETATE; MISMATCH; ALCOHOL;
Keywords:
Saccharomyces cerevisiae; sake yeast; FAS2; point mutation; allele-specific PCR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
22
Recensione:
Indirizzi per estratti:
Indirizzo: Akada, R Yamaguchi Univ, Fac Engn, Dept Chem Engn & Appl Chem, Ube, Yamaguchi 7558611, Japan Yamaguchi Univ Ube Yamaguchi Japan 7558611 aguchi 7558611, Japan
Citazione:
R. Akada et al., "Detection of a point mutation in FAS2 gene of sake yeast strains by allele-specific PCR amplification", J BIOSCI BI, 92(2), 2001, pp. 189-192

Abstract

To identify yeast mutants with a point mutation, detection of the specificmutant alleles is necessary. For this purpose, we applied allele-specifie polymerase chain reaction (PCR) to detect the FAS2-1250S dominant mutant allele that encodes an altered fatty acid synthase in Japanese brewer's yeaststrains. These strains are known to produce a higher amount of ethyl caproate in Japanese sake. The mutant strains were supposed to be diploid and tocontain 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 havean 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 usedsake yeast strains. The method presented here will be useful for the identification of specific yeast strains.

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Documento generato il 01/04/20 alle ore 23:36:59