Construction of a Baculovirus Hyphantria cunea NPV Insecticide Containing the Insecticidal Protein Gene of Bacillus thuringiensis subsp. kurstaki HD1

  • Lee, Hyung-Hoan (Department of Biology & Institute for Genetic Engineering, Konkuk University) ;
  • Moon, Eui-Sik (Department of Biology & Institute for Genetic Engineering, Konkuk University) ;
  • Lee, Sung-Tae (Department of Biology & Institute for Genetic Engineering, Konkuk University) ;
  • Hwang, Sung-Hei (Department of Biology & Institute for Genetic Engineering, Konkuk University) ;
  • Cha, Soung-Chul (Department of Biology & Institute for Genetic Engineering, Konkuk University) ;
  • Yoo, Kwan-Hee (Department of Biology, Sanggi University)
  • 발행 : 1998.12.01

초록

Baculovirus Hyphantrin. cunea nuclear polyhedrosis virus (HcNPV) insecticide containing the insecticidal protein (ICP) gene from Bacillus thuringiensis subsp. kurstaki HD1 was constructed using a lacZ-HcNPV system. The ICP ($\delta$-endotoxin) gene was placed under the control of the polyhedrin gene promoter of the HcNPV. A polyhedrin-negative virus was derived and named ICP-HcNPV insecticide. Then, the insertion of the ICP gene in the ICP-HcNPV genome was confirmed by Southern hybridization analysis. Polyacrylamide gel electrophoresis (PAGE) analysis of the Spodoptera frugiperda cell extracts infected with the ICP-HcNPV showed that the ICP was expressed in the insect cells as 130 kDa at 5 days post-infection. The ICP produced in the cells was present in aggregates. When extracts from the cells infected with the ICP-HcNPV were fed to 20 Bombyx mori larvae, the following mortality rate was seen; 8 larvae at 1 h, 10 larvae at 3 h, and 20 larvae at 12 h. These data indicate that the B. thuringiensis ICP gene was expressed by the baculovirus insecticide in insect cells and there was a high insecticidal activity. The biological activities of the recombinant virus ICP-HcNPV were assessed in conventional bioassay tests by feeding virus particles and ICP to the insect larvae. The initial baculovirus insecticide ICP-HcNPV was developed in our laboratory and the significance of the genetically engineered virus insecticides is discussed.

키워드