Yeast Promoters and lacZ Fusions Designed to Study Expression of Cloned Genes in Yeast

Abstract

This chapter describes plasmids and methods for constructing fusions of any cloned gene to lacZ for study in yeast. If the gene to be fused to lacZ contains no appropriate Sau3A sites, alternative strategies to the above must be employed. If the DNA sequence of the gene to be fused to lacZ is known, inframe fusions can be made simply by choosing the appropriate gene fragment. The vector sequences that precede the yeast DNA is a yeast gene–lacZ fusion may affect gene regulation. Plasmids may be used to probe the signals that govern the initiation of transcription and translation in S. cerevisiae. These methods provide powerful tools in the analysis of the expression of yeast genes. Further, the plasmids should facilitate the expression in yeast of any cloned gene to produce the native, unfused product.