| Literature DB >> 24182917 |
Megan Bergkessel1, Christine Guthrie, John Abelson.
Abstract
It is often useful to replace a small region of the yeast genome containing a gene of interest with a selectable marker. The selectable marker allows for easy identification of yeast cells that have successfully carried out the gene replacement, and functional consequences of the loss of that gene can then be assessed. The same technique can also be used for removing noncoding portions of the genome that may be of interest, such as promoters or 3' or 5' UTRs, and for introducing tags on the N- or C-termini of proteins (alternatively, see a marker-free yeast gene replacement method on Gene Knockouts, in vivo site-directed mutagenesis and Other Modifications Using the Delitto Perfetto System in Saccharomyces cerevisiae).Entities:
Keywords: Agarose gel visualization; Colonies growing on selective media; Deoxyribonucleic acid (DNA); Polyacrylamide gel electrophoresis (PAGE); Polymerase chain reaction (PCR); Transforming yeast with linear PCR product; Yeast-gene replacement
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Year: 2013 PMID: 24182917 DOI: 10.1016/B978-0-12-420067-8.00005-2
Source DB: PubMed Journal: Methods Enzymol ISSN: 0076-6879 Impact factor: 1.600