Modified baculovirus system for high expression of Bombyx mori bidensovirus NS1 in silkworm.
- Author:
Guohui LI
;
Mangmang LI
;
Qian ZHOU
;
Zhaoyang HU
;
Qi TANG
;
Qin YAO
- Publication Type:Journal Article
- MeSH:
Animals;
Baculoviridae;
Bombyx;
virology;
Cell Line;
Chitinases;
Cysteine Proteases;
Genetic Vectors;
Insect Viruses;
Promoter Regions, Genetic;
Transfection
- From:
Chinese Journal of Biotechnology
2015;31(4):591-602
- CountryChina
- Language:Chinese
-
Abstract:
To improve the expression of heterologous genes using baculovirus expression system, we constructed a novel shuttle vector based on the Bm-Bacmid. In the Bm-Bacmid, partial sequences of Chitinase and Cystein Protease were replaced with a tandem cassette of Cm and egfp through homologous recombination. Bombyx mori bidensovirus (BmBDV) ns1 under the control of polyhedrin promoter was inserted into the modified Bm-bacmid by transposition. For comparison, BmBDV ns1 under the control of polyhedrin promoter was also cloned in the wild type Bm-bacmid. The resulting Bm-bacmids were transfected into the cultured BmN cells to prepare recombinant virus to infect silkworms for expression of BmBDV ns1. Total proteins of hemocyte from infected silkworms were subjected to Western blotting and ELISA analysis. The yield of BmBDV NS1 1 with the modified vector was three times as much as that with the unmodified vector. The method to improve the yield of BmBDV NS1 in silkworms will facilitate the function and three-dimensional structure study of BmBDV NS1.
