Yeast artificial chromosome (YAC) vectors permit the cloning of up to megabase fragments of human genomic DNA in yeast (Saccharomyces cerevisiae). Efficient recombination between homologous segments of DNA is one of the hallmark genetic features of yeast. This characteristic facilitates the introduction of specific mutations into YACs by gene targeting. Gene targeting has been used recently to introduce specific mutations into YACs spanning the human β-globin locus and the human apolipoprotein (apo)-B gene, and the mutated YAC DNA has been used to generate transgenic mice. Tiffs approach has been useful for the study of the regulatory elements controlling β-globin gene expression and/or the study of apo-B structure and function. This article reviews the techniques for introducing mutations into YACs, for the purpose of expression in transgenic mice.
ASJC Scopus subject areas
- Cardiology and Cardiovascular Medicine