Random mutagenesis of a gene and subsequent screening for phenotypic properties is a
basic genetic procedure for studying gene function. When working with yeast, cloning …

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) …

This unit describes several procedures for manipulating plasmids in yeast cells. The first is a
general method to segregate autonomously replicating plasmids from cells: plasmid …

The completion of the DNA sequence of the budding yeast Saccharomyces cerevisiae
resulted in the identification of a large number of genes. However, the function of most of …

A major advantage of working with yeast is the ability to replace the wild‐type chromosomal
copy of a gene with a mutant derivative that is constructed in vitro using a cloned copy of the …

Publisher Summary The genetic analysis of essential genes still relies largely on the
generation of point mutations in those genes for an alternative approach, exploiting …

Background A valuable weapon in the arsenal available to yeast geneticists is the ability to
introduce specific mutations into yeast genome. In particular, methods have been developed …

We have adapted a method for making libraries of mutations in any specific gene for use in
the fission yeast Schizosaccharomyces pombe. This elegant and simple method consists of …

We describe here the generation of gene disruption constructs using PCR amplification of
selectable markers with primers that provide homology to the target gene of interest. We find …

Publisher Summary This chapter presents the principles of three new experimental
approaches that aid the analysis of yeast gene function and regulation in vivo by making it …