Genetic Screening using High-Throughput Barcode Sequencing of Saccharomyces cerevisiae Knockout Collection: Search for Regulators of mtDNA Intracellular Selection
A. A. Burlaka, E. S. Glagoleva, N. D. Kashko, D. A. Knorre
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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.
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Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance.
Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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