{"title":"A simple genotyping method for <i>RIM15</i> gene polymorphisms in sake yeast strains.","authors":"Takashi Kuribayashi, Nanako Asahi, Hina Satone, Jumpei Tanaka, Masamichi Sugawara, Keigo Sato, Yoshihito Nabekura, Toshio Joh, Toshio Aoki","doi":"10.47371/mycosci.2024.12.001","DOIUrl":null,"url":null,"abstract":"<p><p>In the process of brewing sake, <i>Saccharomyces cerevisiae</i> produces alcohol and a wide range of flavor compounds. A previous study identified a sake yeast mutant with a single-nucleotide insertion in the <i>RIM15</i> protein kinase gene (<i>RIM15</i> with an insertion of adenine at position 5067: <i>RIM15</i>ins5067A). In this study, we developed primer sets (P1/P2) with engineered mismatches that introduce restriction sites for the restriction enzyme CspCI. This allowed us to create a simple and rapid polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay to identify Kyokai no. 7 (K7) group sake yeast strains carrying the <i>RIM15</i>ins5067A mutation. We validated the PCR-RFLP method in detecting the <i>RIM15</i>ins5067A mutation in the K7 group of yeast strains. This approach provides a straightforward, effective, and valuable tool for distinguishing specific yeasts used in sake production.</p>","PeriodicalId":18780,"journal":{"name":"Mycoscience","volume":"66 2","pages":"134-137"},"PeriodicalIF":1.4000,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13062960/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mycoscience","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.47371/mycosci.2024.12.001","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q4","JCRName":"MYCOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
In the process of brewing sake, Saccharomyces cerevisiae produces alcohol and a wide range of flavor compounds. A previous study identified a sake yeast mutant with a single-nucleotide insertion in the RIM15 protein kinase gene (RIM15 with an insertion of adenine at position 5067: RIM15ins5067A). In this study, we developed primer sets (P1/P2) with engineered mismatches that introduce restriction sites for the restriction enzyme CspCI. This allowed us to create a simple and rapid polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay to identify Kyokai no. 7 (K7) group sake yeast strains carrying the RIM15ins5067A mutation. We validated the PCR-RFLP method in detecting the RIM15ins5067A mutation in the K7 group of yeast strains. This approach provides a straightforward, effective, and valuable tool for distinguishing specific yeasts used in sake production.
期刊介绍：
Mycoscience is the official English-language journal of the Mycological Society of Japan and is issued bimonthly. Mycoscience publishes original research articles and reviews on various topics related to fungi including yeasts and other organisms that have traditionally been studied by mycologists. The research areas covered by Mycoscience extend from such purely scientific fields as systematics, evolution, phylogeny, morphology, ecology, physiology, biochemistry, genetics, and molecular biology, to agricultural, medical, and industrial applications. New and improved applications of well-established mycological techniques and methods are also covered.
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