- Yeast의 FLP/FRT 시스템을 이용한 BmNPV의 유전자 재조합
- ㆍ 저자명
- 강석우,윤은영
- ㆍ 간행물명
- 韓國蠶絲學會誌
- ㆍ 권/호정보
- 1998년|40권 1호|pp.52-59 (8 pages)
- ㆍ 발행정보
- 한국잠사학회
- ㆍ 파일정보
- 정기간행물| PDF텍스트
- ㆍ 주제분야
- 기타
For the construction of plasmid and bmNPV sarrying the FRT recognition site for the FLP recombinases, we synthesized the wild type FRT dligonucleotides. The target FRT sequences consist of three 13bp repeated DNA sequences; two repeats in a direct orientation and one inverted relative to the other two. In addition, there is an 8bp spacer region between the repeats which determune the orientation of the FRT recombination site. In order to place the FRT site both in target BmNPV genome and the transfer vector, we constructed a plasmid, FRT site both in the target BmNPv genome and the transfer vector, we constructed a plasmid, pFRT$eta$-gal, carrying the FRT sites within the cloning sites of pSV vector and a recombinant BmNPV, vFRTPH, carrying the FRT sites at a downstream of polyhedrin promotor, respectively. In order to test the functionality of the FLP/FRT site-specific recombination system, vFRTPH, pFRT$eta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$eta$2-gal. From construction analysis of the vFRT$eta$2-gal with PCR technique it was concluded that the entire pFRT$eta$-gal plasmid with $eta$-galactosidase gene and origines of replication flanked by two functional hybrid FRT sequences. The efficiency of recombination was 8.7%, which was higher than that(2.2%) of recombination between a conventional transfer vector and the wild type BmNPV.