Journal of Microbiology and Biotechnology
- Volume 8 Issue 6
- /
- Pages.676-684
- /
- 1998
- /
- 1017-7825(pISSN)
- /
- 1738-8872(eISSN)
Construction of a Baculovirus Expression System Using Hyphantria cunea Nuclear Polyhedrosis Virus for Eukaryotic Cells
- Lee, Hyung-Hoan (Department of Biology and Institute for Genetic Engineering, Konkuk University) ;
- Kang, Bong-Joo (Department of Biology and Institute for Genetic Engineering, Konkuk University) ;
- Park, Kap-Ju (Department of Biology and Institute for Genetic Engineering, Konkuk University) ;
- Cha, Soung-Chul (Department of Biology and Institute for Genetic Engineering, Konkuk University)
- Published : 1998.12.01
Abstract
Baculovirus transfer and expression vectors with Hyphantria cunea nuclear polyhedrosis virus (HcNPV) were constructed. An initial transfer vector, pHcEV, constructed using HcNPV was previously reported (Park et al. 1993. J. Kor. Soc. Viral. 23: 141-151). Herein, the size of the vector was properly reduced, and a functionally perfect vector was constructed and named pHcEV-IV (6.7 kb). The vector has a 2.2-kb HcNPV DNA sequence in the 5'-flanking region of the vector's polyhedrin gene promoter. The 1.8-kb HcNPV DNA sequence, poly A signal sequence, T3 primer sequence, and 13 multicloning site sequences, in order, were ligated in front of the translation start codon of the polyhedrin gene. The cloning indicating marker lacZ gene was inserted into the pHcEV-IV, named pHcEV-IV-lacZ, and transferred into the wild-type virus. Recombinant expression virus, lacZ-HcNPV, was constructed by replacing the lacZ gene in the pHcEV-IV-lacZ with the polyhedrin gene of the wild-type virus. The recombinant virus was isolated from blue plaques that produce
Keywords
- Baculovirus;
- Hyphantria cunea nuclear polyhedrosis virus;
- cloning and expression vector;
- recombinant virus;
- Spodoptera frugiperda cell;
- lacZ