新城疫病毒F蛋白中两段七肽重复序列的克隆和表达

Cloning and Expression of Heptad Repeat Regions of the Newcastle Disease Virus Fusion Protein

从新城疫病毒 (NDV)中国强毒株F4 8E9和弱毒株长春株F蛋白的cDNA中亚克隆出两段七肽重复序列(HeptadRepeatRegion ,HR1,HR2 ) ,将HR1和HR2分别插入表达载体pGEX 6p 1,在大肠杆菌BL2 1(DE3 )中表达 ,将与载体中的GST(GlutathioneS Trasferase)融合表达的可溶性融合蛋白用GST亲和层析柱纯化。纯化的融合蛋白用蛋白酶酶切后 ,先用GST亲和层析柱除去GST ,再加热进一步纯化。纯化的HR1和HR2质谱分析其分子量 ,结果表明 ,强株的HR1和HR2的分子量分别为 7 10 3kD和 6 3 0 1kD ,弱株的HR1和HR2的分子量分别为 7 10 7kD和6 3 0 9kD ,强弱株HR1和HR2的分子量都基本一致。本工作为研究HR1、HR2的结构以及它们在NDV与宿主细胞融合中的作用奠定了基础。

Two heptad repeat regions (HR1,HR2) from F clone of the virulent and avirulent NDV were cloned,expressed with vector pGEX-6p-1 in %E.coli% BL21 (DE3), and purified the fusion protein by Glutathiones Sepharose 4B Column.After cleaved by prescission protease,the desired protein was purified by Glutathione Sepharose 4B Column and high temperature.Purified HR1 and HR2 were analyzed by Mass spectrum,the result shows that the Molecular Weights of HR1 and HR2 of virulent NDV are 7103 and 6301,and the Molecular Weights of HR1 and HR2 of avirulent NDV is 7107 and 6309.