A. A. Burlaka, E. S. Glagoleva, N. D. Kashko, D. A. Knorre
{"title":"利用高通量条形码测序对酿酒酵母剔除集进行遗传筛选:寻找mtDNA胞内选择的调控因子","authors":"A. A. Burlaka, E. S. Glagoleva, N. D. Kashko, D. A. Knorre","doi":"10.1134/s0026261723604001","DOIUrl":null,"url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>We searched for genes whose deletion affects mitochondrial heteroplasmy in the yeast <i>Saccharomyces cerevisiae</i>. In order to do this, we crossed a yeast knockout collection pool (<i>rho</i><sup><i>+</i></sup>) with a <i>rho</i><sup><i>–</i></sup> strain containing a large deletion in its mtDNA. The resulting mixture of cells was plated onto selective media that made it possible to distinguish between diploid cells that had lost and retained wild-type mtDNA as a result of such crossing. From the resulting pools (initial one, cells that lose <i>rho</i><sup><i>+</i></sup> mtDNA, and those that retained it), we isolated DNA and determined the proportion of deletion strains using high-throughput sequencing. This made it possible to identify functional groups of genes that reduce the ability of yeast cells to maintain <i>rho</i><sup><i>+</i></sup> mtDNA in a state of heteroplasmy with <i>rho</i><sup><i>–</i></sup> mtDNA.</p>","PeriodicalId":18514,"journal":{"name":"Microbiology","volume":"254 1","pages":""},"PeriodicalIF":1.3000,"publicationDate":"2024-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Genetic Screening using High-Throughput Barcode Sequencing of Saccharomyces cerevisiae Knockout Collection: Search for Regulators of mtDNA Intracellular Selection\",\"authors\":\"A. A. Burlaka, E. S. Glagoleva, N. D. Kashko, D. A. Knorre\",\"doi\":\"10.1134/s0026261723604001\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<h3 data-test=\\\"abstract-sub-heading\\\">Abstract</h3><p>We searched for genes whose deletion affects mitochondrial heteroplasmy in the yeast <i>Saccharomyces cerevisiae</i>. In order to do this, we crossed a yeast knockout collection pool (<i>rho</i><sup><i>+</i></sup>) with a <i>rho</i><sup><i>–</i></sup> strain containing a large deletion in its mtDNA. The resulting mixture of cells was plated onto selective media that made it possible to distinguish between diploid cells that had lost and retained wild-type mtDNA as a result of such crossing. From the resulting pools (initial one, cells that lose <i>rho</i><sup><i>+</i></sup> mtDNA, and those that retained it), we isolated DNA and determined the proportion of deletion strains using high-throughput sequencing. This made it possible to identify functional groups of genes that reduce the ability of yeast cells to maintain <i>rho</i><sup><i>+</i></sup> mtDNA in a state of heteroplasmy with <i>rho</i><sup><i>–</i></sup> mtDNA.</p>\",\"PeriodicalId\":18514,\"journal\":{\"name\":\"Microbiology\",\"volume\":\"254 1\",\"pages\":\"\"},\"PeriodicalIF\":1.3000,\"publicationDate\":\"2024-02-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1134/s0026261723604001\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1134/s0026261723604001","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Genetic Screening using High-Throughput Barcode Sequencing of Saccharomyces cerevisiae Knockout Collection: Search for Regulators of mtDNA Intracellular Selection
Abstract
We searched for genes whose deletion affects mitochondrial heteroplasmy in the yeast Saccharomyces cerevisiae. In order to do this, we crossed a yeast knockout collection pool (rho+) with a rho– strain containing a large deletion in its mtDNA. The resulting mixture of cells was plated onto selective media that made it possible to distinguish between diploid cells that had lost and retained wild-type mtDNA as a result of such crossing. From the resulting pools (initial one, cells that lose rho+ mtDNA, and those that retained it), we isolated DNA and determined the proportion of deletion strains using high-throughput sequencing. This made it possible to identify functional groups of genes that reduce the ability of yeast cells to maintain rho+ mtDNA in a state of heteroplasmy with rho– mtDNA.
期刊介绍:
Microbiology is an is an international peer reviewed journal that covers a wide range of problems in the areas of fundamental and applied microbiology. The journal publishes experimental and theoretical papers, reviews on modern trends in different fields of microbiological science, and short communications with descriptions of unusual observations. The journal welcomes manuscripts from all countries in the English or Russian language.