Chromosome engineering in Saccharomyces cerevisiae by using a site-specific recombination system of a yeast plasmid.
We have developed an effective method to delete or invert a chromosomal segment and to create reciprocal recombination between two nonhomologous chromosomes in Saccharomyces cerevisiae, using the site-specific recombination system of pSR1, a circular cryptic DNA plasmid resembling 2 microns DNA of S. cerevisiae but originating from another yeast, Zygosaccharomyces rouxii. A 2.1-kilobase-pair DNA fragment bearing the specific recombination site on the inverted repeats of pSR1 was inserted at target sites on a single or two different chromosomes of S. cerevisiae by using integrative vectors. The cells were then transformed with a plasmid bearing the R gene of pSR1, which encodes the site-specific recombination enzyme and is placed downstream of the GAL1 promoter. When the transformants were cultivated in galactose medium, the recombination enzyme produced by expression of the R gene created the modified chromosome(s) by recombination between two specific recombination sites inserted on the chromosome(s).
Department of Fermentation Technology, Faculty of Engineering, Osaka University, Japan.
H
H
Matsuzaki
R
R
Nakajima
J
J
Nishiyama
H
H
Araki
Y
Y
Oshima
D001483:Base Sequence; D002872:Chromosome Deletion; D007446:Chromosome Inversion; D015825:Chromosomes, Fungal; D005818:Genetic Engineering / Q000379:methods*; D006238:Haploidy; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D011995:Recombination, Genetic; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D015345:Oligonucleotide Probes