Production of recombinant Bombyx mori nucleopolyhedrovirus in silkworm by intrahaemocoelic injection with invasive diaminopimelate auxotrophic Escherichia coli containing BmNPV-Bacmid
文献类型： 外文期刊
作者： Sun, Jingchen 1 ; Yao, Lunguang 1 ; Yao, Ning 3 ; Xu, Hua 1 ; Jin, Pengfei 1 ; Kan, Yunchao 1 ;
作者机构： 1.Nan Yang Normal Univ, Hen Nan Prov Key Lab Funiu Mt Insect Biol, Rothamsted Res Joint Lab Insect Biol, Nan Yang 473064, Peoples R China
2.S China Agr Univ, Coll Anim Sci, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangzhou 510642, Guangdong, Peoples R China
3.SW Jiaotong Univ, Sch Life Sci & Engn, Chengdu 610031, Peoples R China
关键词： Cost-effective and transfectant-free method;Diaminopimelate auxotrophy;Invasion factor;Recombinant Bombyx mori nucleopolyhedrovirus;Silkworm bioreactor
期刊名称：BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY （ 影响因子：2.431； 五年影响因子：2.124 ）
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收录情况： SCI
摘要： The present study elaborates a cost-effective and transfectant-free method for generating recombinant Bombyx mori (silkworm) nucleopolyhedrovirus in silkworm larvae and pupae by injecting invasive Escherichia coli carrying BmBacmid [BmNPV (B. mori nucleopolyhedrovirus)-Bacmid] into larval haemocoel. Up to 109 PFU (plaque-forming units)/ml of infective recombinant baculovirus was generated in the silkworm by intrahaemocoelic injection with 106 DAP (diaminopimelic acid) auxotrophic and BmBacmid containing E. coli cells expressing both invasin and listeriolysin. Thus 1 ml of overnight culture of E. coli is sufficient to inject more than 2000 larvae, while DAP costing up to $1 is enough to inject about 4000 larvae. Recombinant proteins can be controlled to be expressedmainly in pupae by adjusting the injection dose, too. In this new method, many original manipulations have been eliminated, including BmBacmid preparation and the subsequent complex transfection procedures. Hence it is a time- and costsaving means for large-scale injection of B. mori for recombinant baculovirus production in comparison with the traditional transfection methods, which may play an important role in the industrial development of the BmNPV-silkworm bioreactor.
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