水痘-带状疱疹病毒糖蛋白E的纯化及鉴定

Purification and Identification of Recombinant Varicella-Zoster Virus Glycoprotein E

目的 纯化及鉴定重组表达的水痘 -带状疱疹病毒 (VZV)糖蛋白 E(g E)。方法 用 IPTG诱导重组载体 p GEX- VZVg E表达融合蛋白 ,并用亲和层析纯化融合蛋白 ;然后用凝血酶酶切融合蛋白 ,并用免疫印迹法检测酶切产物的抗原性。结果 p GEX- VZVg E的 IPTG诱导表达产物用亲和层析柱去除杂蛋白后 ,获得相对分子质量为 98× 10 3的纯化融合蛋白 ,经凝血酶酶切后得到了相对分子质量大约为 72× 10 3的糖蛋白 E;纯化的融合蛋白和糖蛋白 E均为 SDS- PAGE单点纯 ,并用免疫印迹证明糖蛋白 E具有良好的抗原性。结论 采用亲和层析柱可以有效的纯化重组 VZV糖蛋白 E,从而为 VZV糖蛋白 E的应用研究打下了基础。

Objective To purify and identify recombinant Varicella-Zoster Virus Glycoprotein E. Methods The recombinant plasmid pGEX-VZVgE was induced by isopropy-β-D-thiogalactoside(IPTG), the fusion protein was purified with affinity chromatography column; then the purified fusion protein was cleaved by thrombin, and the product's antigenicity was examined by Western blot. Results The product of pGEX-VZVgE induced by IPTG was separated from the mixture proteins by the affinity chromatography column, the expressed fusion protein's relative molecular mass was about 98×10 3. After cleavage, the obtained VZV Glycoprotein E's relative molecular mass was about 72×10 3; the purified fusion protein and VZV Glycoprotein E were single band by SDS-PAGE; The available antigenicity of Glycoprotein E was confirmed by Western blot.Conclusion Purification of VZV Glycoprotein E with affinity chromatography is an effective method. It provides a foreground for studies on the application of VZV gE.