Isolation of p10 gene fromBombyx mori nuclear polyhedrosis virus and study of its promoter activity in recombinant baculovirus vector system |
| |
Authors: | Shuichiro Tomita Toshimichi Kanaya Jun Kobayashi and Shigeo Imanishi |
| |
Institution: | (1) Department of Insect Genetics and Breeding, National Institute of Sericultural and Entomological Science, Tsukuba, 305 Ibaraki, Japan;(2) Research Institute for Biological Science, Katakura Industries Co., Ltd., Matsumoto, 390 Nagano, Japan;(3) Department of Chemistry for Materials, Faculty of Engineering, Mie University, 1515 Kamihama-cho, 514 Tsu, Mie, Japan |
| |
Abstract: | A homologue ofAutographa californica NPV (AcNPV) p10 gene was identified and cloned fromBombyx mori NPV (BmNPV). BmNPV p10 gene encodes truncated protein of 70 amino acid residues that lacks carboxyl terminus comparing with the p10 protein encoded by AcNPV. The putative TATA box sequence and the ATAAG motif which is the consensus sequence of baculovirus very late promoter were conserved. A transfer vector, pBNT1, which includes the p10 promoter region of BmNPV for foreign gene expression was constructed. By using pBNT1, a recombinant BmNPV, Bmp10-Luc, in which the p10 gene was replaced by the firefly luciferase gene, was obtained. We also obtained another recombinant virus, BmPH-Luc, in which the polyhedrin gene was replaced by the luciferase gene. The luciferase activity detected in BoMo-15AIIc insect cells infected with Bmp10-Luc was approximately 50% of that infected with BmPH-Luc, suggesting that although both the p10 and polyhedrin promoters of BrnNPV are effective in high-level expression of foreign gene, the p10 promoter is not so strong as the polyhedrin promoter. |
| |
Keywords: | baculovirus Bombyx mori expression vector insect cell line promoter activity p10 |
本文献已被 SpringerLink 等数据库收录! |
|