Markers, Selection, and Media in Yeast Artificial Chromosome Cloning

Yeast have proven to be a valuable and flexible host for cloning and manipulating large fragments of foreign genetic material, providing valuable reagents for the study of gene function and regulation in a variety of applications. However, this approach requires a familiarity with basic procedures derived from yeast genetics, and lack of experience can be a barrier to researchers with backgrounds limited to bacterial and mammalian culture systems. To address this issue, this chapter introduces and discusses the genetic markers and selection systems used in the genetic manipulation of yeast, with particular reference to yeast artificial chromosome (YAC) cloning and manipulation. Detailed descriptions of culture media are provided, allowing for the growth and selection of standard YAC strains, as well as for specialist applications.