马传染性贫血病病毒核心蛋白基因的克隆和表达

Expression of Equine Infectious Anemia Virus Core Proteins by Recombinant Baculovirus System

根据美国马传染性贫血病病毒（EIAV）Wyoming株基因序列，设计扩增EIAVgag和p26基因的引物，用PCR法能忠实的分别扩增出全长gag和p26基因。经用杆状病毒表达系统对所获gag和p26基因进行克隆和重组，获得了高效稳定表达Gag和p26蛋白的重组杆状病毒（rBVs）。含有gag和p26基因的重组杆状病毒感染昆虫Sf21细胞，经无血清昆虫细胞培养基增殖和纯化，每升培养物能获得2mgCag或12mgp26蛋白。

In the present studies, we employed the PCR amplification technique to obtain the gag and p26 genes from infected horse macrophage. PCR products not only for the gag but also the p26 genes could be faithfully amplified using the primer sets according to the Sequence of the EIAV Wyoming strain. Tile EIAV core proteins, gag precursor and p26 was expressed by recombinant baculovirus system. The recombinant baculoviuses (rBVs) could highly expressed gag and p26 proteins in Se21 cells with serum free medium. Approximately 2 mg of gag and 12 mg of p26 proteins were obtained from 1 liter of rBV-infected Sf21 cell cultures respectively.