title: Yeast-gene replacement using PCR products.
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).
authors.affiliation: Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: Bergkessel
authors.forename: Christine
authors.initials: C
authors.lastname: Guthrie
authors.forename: John
authors.initials: J
authors.lastname: Abelson
mesh_terms: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel / Q000379:methods; D055786:Gene Knockout Techniques / Q000379:methods*; D005819:Genetic Markers; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015003:Yeasts / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D017931:DNA Primers; D005819:Genetic Markers
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