• Title/Summary/Keyword: SV 40 replication system

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Inhibitory Effects of Extracts of Paralichtys olivaceus on DNA replication at the Level of Initiation (Paralichthys olivaceous 추출물에 의한 CNA 복제 개시 단계에서의 억제 효과)

  • 이지현;임영해;이수복;김동규;김동선;박남규;정준기;김남득
    • Journal of Life Science
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    • v.11 no.4
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    • pp.371-378
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    • 2001
  • The effects of the RM 60 series on DNA replication systems were examined by using simian virus 40 (SV40) DNA replication system in vitro. The RM 60 series inhibited the DNA replication in the initiation step rather than in the elongation step. Polymerase$\alpha$-primase activity and toposiomerase I activity study were performed subsequently, the RM 60 seies increased the polymerase $\alpha$-primase activity in a low dose, but decreased the activity in a higher dose. The topoisomerase I activity was inhibited by the RM 60 series. This result suggests that the RM 60 series might inhibit some molecules which are required to establish replication forks during the initiation step of the DNA replication.

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Construction of Recombinant Bombyx mori Nuclear Polyhedrosis Virus Using a FLP/FRT System of Yeast, Saccharomyces cerevisiae 2$\mu$m plasmid (Yeast의 FLP/FRT 시스템을 이용한 BmNPV의 유전자 재조합)

  • 강석우;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.40 no.1
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    • pp.52-59
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    • 1998
  • 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$\beta$-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$\beta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$\beta$2-gal. From construction analysis of the vFRT$\beta$2-gal with PCR technique it was concluded that the entire pFRT$\beta$-gal plasmid with $\beta$-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.

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