A rapid method for localized mutagenesis of yeast genes - PubMed (original) (raw)
A rapid method for localized mutagenesis of yeast genes
D Muhlrad et al. Yeast. 1992 Feb.
Abstract
We have developed a simple procedure for the localized mutagenesis of yeast genes. In this technique the region of interest is first amplified under mutagenic polymerase chain reaction (PCR) conditions. Cotransformation of the PCR product with a gapped plasmid containing homology to both ends of the PCR product allows in vivo recombination to repair the gap with the mutagenized DNA. This procedure is efficient, allows targeting of specific regions for mutagenesis, and requires no subcloning steps in Escherichia coli.
Similar articles
- In vivo site-directed mutagenesis using oligonucleotides.
Storici F, Lewis LK, Resnick MA. Storici F, et al. Nat Biotechnol. 2001 Aug;19(8):773-6. doi: 10.1038/90837. Nat Biotechnol. 2001. PMID: 11479573 - Recombination-mediated PCR-directed plasmid construction in vivo in yeast.
Oldenburg KR, Vo KT, Michaelis S, Paddon C. Oldenburg KR, et al. Nucleic Acids Res. 1997 Jan 15;25(2):451-2. doi: 10.1093/nar/25.2.451. Nucleic Acids Res. 1997. PMID: 9016579 Free PMC article. - Gene knockouts, in vivo site-directed mutagenesis and other modifications using the delitto perfetto system in Saccharomyces cerevisiae.
Stuckey S, Storici F. Stuckey S, et al. Methods Enzymol. 2013;533:103-31. doi: 10.1016/B978-0-12-420067-8.00008-8. Methods Enzymol. 2013. PMID: 24182920 - Rapid assessment of S. cerevisiae mating type by PCR.
Huxley C, Green ED, Dunham I. Huxley C, et al. Trends Genet. 1990 Aug;6(8):236. doi: 10.1016/0168-9525(90)90190-h. Trends Genet. 1990. PMID: 2238077 No abstract available. - Recombinant circle PCR and recombination PCR for site-specific mutagenesis without PCR product purification.
Jones DH, Winistorfer SC. Jones DH, et al. Biotechniques. 1992 Apr;12(4):528-30, 532, 534-5. Biotechniques. 1992. PMID: 1503755
Cited by
- Shuttle vectors for facile gap repair cloning and integration into a neutral locus in Candida albicans.
Gerami-Nejad M, Zacchi LF, McClellan M, Matter K, Berman J. Gerami-Nejad M, et al. Microbiology (Reading). 2013 Mar;159(Pt 3):565-579. doi: 10.1099/mic.0.064097-0. Epub 2013 Jan 10. Microbiology (Reading). 2013. PMID: 23306673 Free PMC article. - Tim54p connects inner membrane assembly and proteolytic pathways in the mitochondrion.
Hwang DK, Claypool SM, Leuenberger D, Tienson HL, Koehler CM. Hwang DK, et al. J Cell Biol. 2007 Sep 24;178(7):1161-75. doi: 10.1083/jcb.200706195. J Cell Biol. 2007. PMID: 17893242 Free PMC article. - Pga1 is an essential component of Glycosylphosphatidylinositol-mannosyltransferase II of Saccharomyces cerevisiae.
Sato K, Noda Y, Yoda K. Sato K, et al. Mol Biol Cell. 2007 Sep;18(9):3472-85. doi: 10.1091/mbc.e07-03-0258. Epub 2007 Jul 5. Mol Biol Cell. 2007. PMID: 17615295 Free PMC article. - Identification of a second transforming function in bovine papillomavirus type 1 E6 and the role of E6 interactions with paxillin, E6BP, and E6AP.
Das K, Bohl J, Vande Pol SB. Das K, et al. J Virol. 2000 Jan;74(2):812-6. doi: 10.1128/jvi.74.2.812-816.2000. J Virol. 2000. PMID: 10623743 Free PMC article. - A role for BiP as an adjustor for the endoplasmic reticulum stress-sensing protein Ire1.
Kimata Y, Oikawa D, Shimizu Y, Ishiwata-Kimata Y, Kohno K. Kimata Y, et al. J Cell Biol. 2004 Nov 8;167(3):445-56. doi: 10.1083/jcb.200405153. Epub 2004 Nov 1. J Cell Biol. 2004. PMID: 15520230 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous