construction of a baculovirus-silkworm multigene expression system and its application on producing virus-like particles基因表达系统及其应用建设baculovirus-silkworm生产病毒样颗粒.pdf

Construction of a Baculovirus-Silkworm Multigene Expression System and Its Application on Producing Virus-Like Particles 1 . 1,2. 2 1,3 4 1,2 1,2 Lunguang Yao * , Shanshan Wang , Shuo Su , Ning Yao , Jian He , Li Peng , Jingchen Sun * 1 China-United Kingdom Nanyang Normal University-Rothamsted Research Joint Laboratory of Insect Biology, Henan Provincial Key Laboratory of Funiu Mountain Insect Biology, Nanyang Normal University, Nanyang, People’s Republic of China, 2 Guangdong Provincial Key Lab of Agro-animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, People’s Republic of China, 3 School of Life Science and Engineering, Southwest Jiaotong University, Chengdu, People’s Republic of China, 4 Guangzhou East Campus Lab Center, Sun Yat-sen University, Guangzhou, People’s Republic of China Abstract A new baculovirus-silkworm multigene expression system named Bombyx mori MultiBac is developed and described here, by which multiple expression cassettes can be introduced into the Bombyx mori nuclear polyhedrosis virus (BmNPV) genome efficiently. The system consists of three donor vectors (pCTdual, pRADM and pUCDMIG) and an invasive diaminopimelate (DAP) auxotrophic recipient E. coli containing BmNPV-Bacmid (BmBacmid) with a homologous recombination region, an attTn7 site and a loxp site. Two genes carried by pCTdual are firstly inserted into BmBacmid by homologous recombination, while the other eight genes in pRADM and pUCDMIG are introduced into BmBacmid through Tn7 transposition and cre-loxp recombination. Then