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

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Bmd3a基因克隆到杆状病毒转移载体pFastBacl中获得了pFastBac-IEl-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24 h和48 h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57 kD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 baculovirus was obtained.The green fluorescence can be observed in the BmN 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 BmN cells with the recombination baculovirus 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.