Cloning of industrial Saccharomyces 2-microns plasmid variants by in vivo site-specific recombination.
Southern analyses defined several industrial Saccharomyces yeast strains with extensive 2-microns DNA polymorphism. Variants included insertions and deletions up to several hundred base pairs. To facilitate the investigation of yeast plasmid evolution we developed a novel method of cloning 2-microns plasmids by taking advantage of 2-microns circle in vivo site-specific recombination and an SMRI gene as a dominant selectable marker. This method can be applied to other organisms for the isolation of plasmid variants and provides a new approach to in vivo plasmid construction.
Xiao
W
W
Department of Biology, University of Saskatchewan, Saskatoon, Canada.
Rank
G H
GH
D003001:Cloning, Molecular; D010957:Plasmids; D012150:Polymorphism, Restriction Fragment Length; D011995:Recombination, Genetic; D012440:Saccharomyces / Q000235:genetics*; D014170:Transformation, Genetic