Development of reusable split URA3-marked knockout vectors for Saccharomyces cerevisiae

초록

Two knockout vectors, in which the truncated Kluyveromyces lactis URA3 gene is flanked by a direct repeat, were developed for Saccharomyces cerevisiae. Each vector was designed to harbor 5'- and 3'-end homology regions for integration. Two knockout fragments were devised to integrate into the correct locus in a complementary manner to disrupt a gene of interest and concomitantly to make functional Kl URA3 for transformant selection. The use of dual complementary knockout cassettes was expected to dramatically reduce integration into unwanted loci in the genome. The knockout system developed in this study was successfully used for disruption of the GAL1 gene in S. cerevisiae.

키워드