Article
Making temperature-sensitive mutants.
Methods in enzymology - 1 Jan 2010
Ben-Aroya Shay, Pan Xuewen, Boeke Jef D, Hieter Philip
Abstract excerpt
The study of temperature-sensitive (Ts) mutant phenotypes is fundamental to gene identification and for dissecting essential gene function. In this chapter, we describe two "shuffling" methods for producing Ts mutants using a combination of PCR, in vivo recombination, and transformation of diploid strains heterozygous for a knockout of the desired mutation. The main difference between the two methods is the type...
Topics
- Alleles
- Diploidy
- Mutation
- Plasmids
- Polymerase Chain Reaction
- Recombination, Genetic
- Saccharomyces cerevisiae
- Temperature
- Transformation, Genetic