A rapid method for localized mutagenesis of yeast genes
- 1 February 1992
- Vol. 8 (2) , 79-82
- https://doi.org/10.1002/yea.320080202
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 chain reaction (PCR) conditions. Cotransformation of the PCR product with a gapped plasmid containing homology to both ends the PCR procuct allows in vivo recombination to repair the gap with the mutagenized DNA. This procedure is efficient, allows trageting of specific regions for mutagenesis, and requires no subcloning steps in Escherichia coli.Keywords
This publication has 10 references indexed in Scilit:
- [20] In vitro mutagenesis and plasmid shuffling: From cloned gene to mutant yeastPublished by Elsevier ,1991
- Multiple roles for U6 snRNA in the splicing pathway.Genes & Development, 1990
- High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrierCurrent Genetics, 1989
- A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiaeGene, 1988
- Plasmid construction by homologous recombination in yeastGene, 1987
- Strategies and Applications of in Vitro MutagenesisScience, 1985
- A General Method for Saturation Mutagenesis of Cloned DNA FragmentsScience, 1985
- Transformation of intact yeast cells treated with alkali cationsJournal of Bacteriology, 1983
- Isolation and genetic analysis of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by a factor and alpha factor pheromones.Molecular and Cellular Biology, 1982