Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Development of Reusable Split URA3-Marked Knockout Vectors for Saccharomyces cerevisiae

  • Lee Tae-Hee (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University) ;
  • Kim Myoung-Dong (School of Biotechnology and Bioengineering, Kangwon National University) ;
  • Seo Jin-Ho (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
  • Published : 2006.06.01
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Abstract

Two knockout vectors, in which the truncated Kluyveromyces lactis URAS 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 transfomant 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.

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References

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