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Efficient and Eco-friendly Recombinant Baculovirus Insecticide

  • 언어ENG
  • URLhttps://db.koreascholar.com/Article/Detail/291264
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한국응용곤충학회 (Korean Society Of Applied Entomology)
초록

A novel recombinant baculovirus, NeuroBactrus, was constructed to develop an improved baculovirus insecticide with additional beneficial properties such as higher insecticidal activity and recovery to wild-type baculovirus. For this, Bacillus thuringiensis crystal protein gene (cry1-5) was introduced into Autographa californica nucleopolyhedrovirus (AcMNPV) genome by fusion of polyhedrincry1- 5-polyhedrin under the control of poyhedrin gene promoter. In the opposite direction of this fusion gene, an insect-specific neurotoxin gene (AaIT) under the control of early promoter from Cotesia plutellae bracovirus was introduced by fusion of orf603 partial fragment. Western hybridization and confocal microscopy revealed that AaIT neurotoxin and Polyhedrin-Cry1-5-Polyhedrin fusion protein expressed by the NeuroBactrus and that the fusion protein occluded into the polyhedra. In addition, the fusion protein was activated as about 65 kDa of crystal protein when treated with trypsin. The NeuroBactrus showed high level of insecticidal activity against Plutella xylostella larvae and significant reduction in median lethal time (LT50) against Spodoptera exigua larvae compared to those of wild-type AcMNPV. Re-recombinants derived from the NeuroBactrus, NBt-Del5 (deleted cry1-5), NBt-DelA (deleted AaIT) and NBt-Del5A (deleted cry1-5 and AaIT; wild-type baculovirus) were generated in serial passages in vitro and in vivo. These results suggested that the NeuroBactrus could be transferred to wild-type baculovirus along with serial passages by the homologous recombination between two polyhedrin genes and two partial orf603 fragments.

저자
  • Hee Jin Shim(Department of Agricultural Biotechnology, Seoul National University)
  • Jae Young Choi(Research Institute for Agriculture and Life Sciences, Seoul National University)
  • Yong Wang(Department of Agricultural Biotechnology, Seoul National University)
  • Jong Yul Roh(Department of Agricultural Biotechnology, Seoul National University)
  • Soo Dong Woo(College of Agriculture, Life and Environment Sciences, Chungbuk National University)
  • Byung Rae Jin(College of Natural Resources and Life Science, Dong-A University)
  • Yeon Ho Je(Department of Agricultural Biotechnology, Seoul National University)