Chromosome bisection in the yeast Saccharomyces cerevisiae facilitated by yeast artificial chromosomes bearing a site-specific recombination system.
A chromosome bisection method was constructed using yeast artificial chromosomes (YAC) and a GAL1-promoted site-specific recombination system. This method was applied to bisect chromosome IV into the left and right parts of the centromere region. The bisection occurred at frequencies of about 10% when the recombination site DNAs were integrated onto YAC and chromosome IV in the same direction, but were less than 10(-3) when they were in opposite directions. Reconstruction of the original chromosome IV from the bisected chromosomes was also induced by galactose at high frequencies. Loss of the left part chromosome was found at the frequencies of 0.9 x 10(-3) after hybrid cells between the chromosome-bisected strain and a normal haploid were subcultivated in a complete medium. The bisection and reconstruction of chromosome IV and deletion of the left part chromosome were demonstrated by electrophoretical karyotyping.
Kawasaki
H
H
Tokyo Research laboratories, Kyowa Hakko Kogyo Co., Ltd., Japan.
Ouchi
K
K
D002872:Chromosome Deletion; D002875:Chromosomes / Q000737:chemistry / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D004171:Diploidy; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D014170:Transformation, Genetic