• 한국잠사곤충학회지
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• 제39권1호
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• pp.36-43
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• 1997

In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

• Journal of Microbiology and Biotechnology
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• 제26권4호
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• pp.725-729
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• 2016

A novel genome-engineering tool in Clostridium acetobutylicum was developed based on single-crossover homologous recombination. A small-sized non-replicable plasmid, pHKO1, was designed for efficient integration into the C. acetobutylicum genome. The integrated pHKO1 plasmid backbone, which included an antibiotic resistance gene, can be excised in vivo by Flp recombinase, leaving a single flippase recognition target sequence in the middle of the targeted gene. Since the pSHL-FLP plasmid, the carrier of the Flp recombinase gene, employed the segregationally unstable pAMβ1 replicon, the plasmid was rapidly cured from the mutant C. acetobutylicum. Consequently, our method makes it easier to engineer C. acetobutylicum.

• 생명과학회지
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• 제17권4호
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• pp.587-592
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• 2007

생쥐 유전자를 과발현 시키거나 제거하는 유전자 조작 기술의 발달은 배 발생 단계나 출생 후 특정한 세포에서의 특정 단계에서의 유전자 기능을 이해하는 많은 기여를 하고 있다. 특히, 높은 상동 재조합 활성을 가지는 생쥐 배 줄기세포를 이용한 유전자 적중 기법은 인간 질환을 이해하는데 필수적인 동물 모델 개발에 중요한 기여를 하였다. 최근에는 Cre과 Flp와 같은 염기서열 특이적 재조합 효소와 라이겐드에 의한 조절 시스템의 도입으로 좀 더 정확하고 정교한 유전자 발현 조절을 위해 개발되어 복잡한 생명현상을 지배하는 메카니즘과 시간과 공간에서 작동하는 유전자의 기능을 이해하는데 많은 기여를 하고 있다. 마우스 게놈을 세밀하게 조작할 수 있는 새로운 분자생물학적 도구의 적용으로 in vivo상에서 유전자의 다양한 기능을 좀 더 정확하게 이해할 수 있는 기회가 열릴 것으로 기대된다.