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Yeast 2 $mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현
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  • Yeast 2 $mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현
저자명
강석우,윤은영
간행물명
韓國蠶絲學會誌
권/호정보
1997년|39권 1호|pp.36-43 (8 pages)
발행정보
한국잠사학회
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정기간행물|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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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 $eta$-galactosidase gene as a reporter gene. The result showed that the pHs $eta$-gal plasmid vector expressed the $eta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $eta$-gal vector, but different from that of a recombinant virus, vBm $eta$-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.