한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제23권5호
  • /
  • Pages.544-549
  • /
  • 1995
  • /
  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

고초균(Bacillus) 염색체상에서 외래 유전자 Alkaline Elastase Gene의 증폭

Multiple Chromosomal Integration of a Bacillus Ya-B Alkaline Elastase Gene

  • 김병문 (한국과학기술연구원 생명공학연구소) ;
  • 정봉현 (한국과학기술연구원 생명공학연구소)
  • 발행 : 1995.10.01
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초록

The alkaline elastase is an extracellular serine protease of the alkalophilic Bacillus strain Ya-B. To increase the gene copy number and the production level of the alkaline elastase Ya-B, we designed, on the B. subtilis chromosome, a gene amplification of the 10.6 kb repeating unit containing amyE, aleE (alkaline elastase Ya-B gene) and tmrB. The aleE was inserted between amyE and tmrB, and B. subtilis APT119 strain was transformed with this amyE-aleE-tmrB-junction region fragment. As a result, we succeeded in obtaining tunicamycin-resistant (Tm$^{r}$) transformants (Tf-1, Tf-2) in which the designed gene amplification of 10.6 kb occurred in chromosome. The transformants showed high productivity of $\alpha $-amylase and alkaline elastase Ya-B. The copy number of the repeating unit (amyE-aleE-tmrB) was estimated to be 25, but plasmid vector (pUC19) was not integrated. The amplified aleE of chromosome was more stable than that of plasmid in absence of antibiotics.

키워드

참고문헌

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