TY - CHAP
T1 - Yeast-gene replacement using PCR products
AU - Bergkessel, Megan
AU - Guthrie, Christine
AU - Abelson, John
PY - 2013
Y1 - 2013
N2 - 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).
AB - 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).
KW - Agarose gel visualization
KW - Colonies growing on selective media
KW - Deoxyribonucleic acid (DNA)
KW - Polyacrylamide gel electrophoresis (PAGE)
KW - Polymerase chain reaction (PCR)
KW - Transforming yeast with linear PCR product
KW - Yeast-gene replacement
UR - http://www.scopus.com/inward/record.url?scp=84887243199&partnerID=8YFLogxK
U2 - 10.1016/B978-0-12-420067-8.00005-2
DO - 10.1016/B978-0-12-420067-8.00005-2
M3 - Chapter
C2 - 24182917
AN - SCOPUS:84887243199
SN - 9780124200678
VL - 533
T3 - Methods in Enzymology
SP - 43
EP - 55
BT - Laboratory Methods in Enzymology
A2 - Lorsch, Jon
PB - Academic Press Inc.
ER -