家蚕Bms3a基因真核表达载体的构建及其在家蚕细胞和蚕体中的表达  

Construction of Bms3a Eukaryotic Expression Vector and Its Expression in BmN Cells and Silkworm,Bombyx mori

作　　者：尤征英[1] 周武松[1] 徐家萍[1] 杨莹[1] 凌琳[1] 王文兵[2] 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036 [2]江苏大学生命科学研究院,江苏镇江212013

出　　处：《激光生物学报》2011年第2期212-218,共7页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(10975002);安徽省科技攻关项目(08010302152)

摘　　要：Bms3a基因可能在家蚕Bombyx mori抗病或细胞凋亡中有一定的作用。将融合有绿色荧光蛋白的Bins3a基因克隆到杆状病毒转移载体pFastBac1中获得了pFastBac-IE1-Bms3a-EGFP真核表达载体,利用杆状病毒(Bac-to-Bac)表达系统筛选重组杆状病毒,以重组病毒感染家蚕BmN细胞和五龄幼虫,分别在感染24h和48h检测到有绿色荧光蛋白的表达,Western blot证明表达的融合蛋白在相对分子量约57kD处出现特异条带,与预计的蛋白理论值相符。结果表明BmS3A-EGFP融合蛋白在家蚕细胞BmN及幼虫体中得到高效表达。研究结果为进一步研究BmS3A蛋白的功能奠定了基础。Bms3a gene may be involved in silkworm resistance and anti-apoptosis. The fragment of Bms3a gene with EGFP has been inserted into a transfer vector pFastBacl and the recombination baeulovirus was obtained. The green fluorescence can be observed in the BraN cells and the 5th instar larvae after 24 h and 48 h by infection of the recombinant virus, respectively. Western blot showed specific reaction band with a relative molecular mass of about 57 kD, which was consistent with predicted size of the target protein. The results showed that the Bms3A protein with EGFP was expressed in BraN cells with the recombination baeulovirus and the Bms3 A protein was highly expressed, which will laid a fundation the basic material for the further mechanistic study of Bms3 A protein.

关 键 词：家蚕 Bins3a基因 真核表达 杆状病毒表达系统 

分 类 号：Q956[生物学—动物学]