细胞适应株狂犬病病毒反向遗传系统辅助质粒的构建

目的 构建细胞适应株狂犬病病毒反向遗传系统中所需的4个结构蛋白辅助质粒.方法 利用RT-PCR方法扩增细胞适应株狂犬病毒的核蛋白(N)、磷酸化蛋白(P)、糖蛋白(G)、转录大蛋白(L)基因,测序结果正确后,分别连接表达载体,构建反向遗传所需的4个功能性蛋白辅助质粒.结果成功扩增4个结构基因,经测序完全正确,并成功构建反向遗传体系中N蛋白、P蛋白、G蛋白、L蛋白的辅助表达质粒,酶切鉴定完全正确.结论 成功构建狂犬病病毒拯救系统的4个辅助质粒,为此细胞适应株狂犬病病毒的拯救奠定基础.

Construction of helper plasmids used in reverse genetic system of a cell adapted rabies virus strain

Objective To construct four helper plasmids which are necessary in reverse genetic system of a cell adapted rabies virus strain. Methods Nucleoprotein gene, phosphprotein gene, glycoprotein gene and the viral RNA polymerse gene are amplified by RT-PCR. The four gene fragments were cloned into an expression vector respectively after sequencing to construct four helper plasmids. Results The four structral genus were amplified successfully, the sequence were correct. And all the structral protein gene fragments were cloned into expression vectors, the results were correct identified by cleavage reaction. Conclusion Construction of the four helper plasmids have been done successfully, and it is the basis for rescuing the adapted cell rabies virus strain from cloned cDNA.