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Construction of recombinant Autographa californica nucleopolyhedrovirus containing genomic segments of Spodoptera litura granulovirus

  • 언어ENG
  • URLhttps://db.koreascholar.com/Article/Detail/290423
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한국응용곤충학회 (Korean Society Of Applied Entomology)
초록

Recently, the genome of Spodoptera litura granulovirus (SlGV) which encodes 133 putative open reading frames (ORFs) was completely sequenced. In this study, to screen novel insecticidal genes of SlGV, we first constructed an advanced plasmid capture system, pPCS-TPI, which contains not only pUC19 ori and ampicillin resistance gene but also Autographa californica nucleopolyhedrovirus (AcMNPV) ORF603 and ORF1629 homologous region between Tn7L and Tn7R. In order to introduce genomic segments of SlGV into the genome of AcMNPV, genomic DNA of SlGV was digested with EcoRI and self-ligated. These self-ligated segments were in vitro transposed with the pPCS-TPI donor by the help of TnsABC* transposase. By this, 10 EcoRI-digested genomic segments of the SlGV were cloned, and these clones were co-transfected with the bApGOZA DNA into sf9 cells to generate corresponding recombinant virus, respectively. The resulting recombinant viruses harboring genomic segments of the SlGV could be used to investigate the insecticidal activity and/or other functions originated from the introduced genomic segments of the SlGV.

저자
  • Yong Wang(Department of Agricultural Biotechnology, Seoul National University)
  • Jae Young Choi(Research Institute for Agriculture and Life Sciences, Seoul National University)
  • Jong Yul Roh(Department of Agricultural Biotechnology, Seoul National University)
  • Hee Jin Shim(Department of Agricultural Biotechnology, Seoul National University)
  • Qin Liu(Department of Agricultural Biotechnology, Seoul National University)
  • Xueying Tao(Department of Agricultural Biotechnology, Seoul National University)
  • Yeon Ho Je(Department of Agricultural Biotechnology, Seoul National University)