Constructing yeast libraries

Abstract

Publisher Summary Having a good library is crucial for gene cloning in Saccharomyces cerevisiae. Different types of libraries are used for different applications. The most frequently used approach in cloning a yeast gene is complementation of a recessive mutant with a yeast genomic library on a low-copy-number yeast vector. Literally, hundreds of yeast genes have been cloned this way. Another popular approach that has efficiently identified many yeast genes is by functional cloning from overexpression libraries, based on the suppression of a recessive mutant by gene overexpression or other phenotypic consequences associated with overexpression. Although numerous yeast genomic libraries are available, many circumstances require the construction of a new library. Methods for de novo construction of yeast genomic libraries and considerations involved in the process have been described in the previous volume in this series. This chapter describes two specific strategies that have been used successfully for the construction of several yeast libraries. One strategy should be applicable to the construction of any genomic library and the other for any cDNA libraries.