Vectors for High-Level, Inducible Expression of Cloned Genes in Yeast

This chapter describes the design and application of vectors that permit high-level, inducible synthesis of the product of a cloned gene in yeast, and why yeast systems should be included in the arsenal of genetic engineers.

Abstract

This chapter describes the design and application of vectors that permit high-level, inducible synthesis of the product of a cloned gene in yeast. These vectors have been developed in recognition of the increasing need for obtaining reasonable amounts of the product of a cloned gene in research projects. For instance, given the variety of cloning strategies currently available, it is often easier to clone DNA encoding a particular gene than to purify the product of that gene. However, appreciation of the biological function of such a gene usually requires the subsequent identification and characterization of its product. There are a number of host-vector systems currently available for the high-level expression of cloned genes. However, there are a number of reasons why yeast systems should be included in the arsenal of genetic engineers. First, the codon bias in yeast is significantly different from that in Escherichia coli, as is, undoubtedly, the spectrum of endogenous proteolytic enzymes. In addition, yeast is capable of promoting glycosylation of newly synthesized proteins.