水疱性口炎病毒糖蛋白基因的真核表达及其初步应用

Eukaryotic expression of vesicular stomatitis virus glycoprotein gene and its primary application

为验证水疱性口炎病毒(vesicular stomatitis virus,VSV)糖蛋白与半乳糖凝集素1(lectin galactoside-binding soluble 1,LGALS1)之间的相互作用,本研究将VSV-G全基因插入至真核表达载体pEF1α-IRES-DsRed-Express2中,构建了真核表达质粒pEF1α-VSV-G。在DNA转染试剂的作用下,将pEF1α-VSV-G转染至BHK-21细胞中,转染后24h经荧光显微镜观察即可见细胞内有红色荧光信号。提取细胞蛋白后经Western-blotting检测,证明VSV-G蛋白成功表达。将表达的蛋白与LGALS1共同用于免疫共沉淀试验,Western-blotting检测结果说明表达的蛋白可以用于蛋白相互作用的研究。本试验为深入开展VSV-G蛋白受体的研究奠定了基础。

To validate the interaction between vesicular stomatitis virus glycoprotein （VSV-G） and lectin galactoside-binding soluble 1 （LGALS1）,VSV-G gene was cloned and inserted into pEFla- IRES-DsRed-Express2 vector to establish pEFla-VSV-G recombinant. After that, the recombi- nant was transfected into BHK-21 cells. VSV-G protein was successfully expressed that were confirmed by Western-blotting and immunofluorescence staining in the transfected cells at 24 hours after the transfection. The result of co-immunoprecipitation experiment indicated that the VSV-G protein can be used in the study of protein interactions. These results will facilitate future research of VSV-G receptors.