Biotechnology and Bioprocess Engineering:BBE

  • 제5권5호
  • /
  • Pages.327-331
  • /
  • 2000
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  • 1226-8372(pISSN)
  • /
  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Application of LFH-PCR for the Disruption of SpoIIIE and SpoIIIG of B. subtilis

  • Kim, June-Hyung (Institute for Molecular Biology and Genetics, and Division of Chemical Enginering, Seoul National University) ;
  • Kim, Byung-Gee (Institute for Molecular Biology and Genetics, and Division of Chemical Enginering, Seoul National University)
  • 발행 : 2000.09.01
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초록

The application of LFH-PCR(long flanking homology region-PCR) for Bacillus subtilis gene disruption is presented. Without plasmid- or phage-vector construction, only by PCR, based on a DNA sequence retrieved from B. subtilis genome data base, kanamycin resistance gene was inserted into two genes of B. subtilis involved in sporulation, spoIIIE and spoIIIG. The effect of gene disruption on subtilisin expression was examined and the sporulation frequency of two mutants was compared to that of the host strain. For this purpose, only 2 or 3 rounds of PCR were required with 4 primers. We first demonstrated the possibility of LFH-PCR for rapid gene disruption to characterize an unknown functional gene of B. subtilis or other prokaryote in the genomic era.

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