Yeast artificial chromosome modification and manipulation

This chapter describes yeast artificial chromosome (YAC) modification and manipulation. Two types of YAC modification systems have been developed that take advantage of this property to target recombination to highly repetitive DNA elements within YAC DNA inserts. Vectors that integrate into YACs can be used to produce chromosome-specific libraries from somatic cell hybrids or to introduce deletions, mutations, or mammalian selectable markers into YAC cloned segments. A second vector system facilitates analysis of the large inserts in YACs by fragmenting them, generating nested deletion derivatives. A polyethylene glycol-mediated fusion procedure has been described that can be used to introduce appropriately modified YACs into cultured mammalian cells. This combination of vectors and procedures provides a system for realizing the full potential of YACs in structural and biological analysis of the genome.