Expression of Bovine Growth Hormone Gene in a Baculovirus, Hyphantria cunea Nuclear Polyhedrosis Virus.
- Author:
Kap Ju PARK
;
Keun Kwang LEE
;
Bong Ju KANG
;
Sung Chul CHA
;
Hyung Hoan LEE
- Publication Type:Original Article
- Keywords:
Baculovirus vector;
Bovine growth hormone;
Hyphantria cunea nuclear polyhedrosis virus
- MeSH:
Baculoviridae*;
Cell Line;
Clone Cells;
DNA;
DNA, Complementary;
Growth Hormone*;
Insects;
Nucleopolyhedrovirus*;
Plasmids;
Polymerase Chain Reaction;
Radioimmunoassay;
Spodoptera
- From:Journal of the Korean Society of Virology
1998;28(2):129-138
- CountryRepublic of Korea
- Language:English
-
Abstract:
Bovine growth hormone (bGH) gene was expressed in an insect spodoptera frugiperda cell line using a Baculovirus, Hyphantria cunea nuclear polyhedrosis virus (HcNPV). The bGH gene in pbGH plasmid was sequenced and amplified by PCR technique with two primers containing NcoI sites. The bGH gene consisted of 654 bp (217 amino acid residues), the 5'-untranslated region of the cloned bGH cDNA contains 56 bp, and the 3'-untranslated region contains 145 bp and two pallindromic regions. The amplified bGH gene DNA fragment (654 bp) was inserted into the NcoI site of the pHcEVII vector, which was named pHcbGH. The pHcbGH transfer vector DNA and the wild type HcNPV DNA were cotransfected into s. frugiperda cells to construct a recombinant virus. Eight recombinant viruses were selected and named HcbGH. One clone, HcbGH-4-1 showed largest plaque size, therefore the recombinant virus was further studied. The multiplication patters of the recombinant HcbGH-4-1 was similar to that of the wild type HcNPV. The bGH gene DNA in the HcbGH-4-1 recombinant was confirmed by Southern lot hybridization. The amount of the bGH (217 amino acid residues, 21 kDa) produced in S. frugiperda cells infected with the HcbGH-4-1 recombinant was approximately 5.5 ng per ml (106 cells) by radioimmunoassay.
