摘要
目的原核表达水痘-带状疱疹病毒(varicella-zoster virus,VZV)糖蛋白E (gE)基因,并对其进行鉴定。方法构建大肠埃希菌重组载体pET28a-gE-his,利用IPTG诱导表达gE蛋白,经亲和层析纯化得到蛋白,用免疫印迹法验证目的蛋白的特异性。结果 pET28a-gE-his/BL21的IPTG诱导表达产物利用亲和层析纯化除去杂蛋白后,获得相对分子质量约为60×10~3的纯化蛋白,并且能与鼠抗gE糖蛋白单抗发生特异性结合,证明该蛋白为gE目的蛋白。结论成功利用大肠埃希菌表达重组VZV糖蛋白E,并利用亲和层析柱进行有效的纯化,从而为VZV糖蛋白E的应用研究打下了基础。
Objective To express and identify the glycoprotein E（gE）of varicella-zoster virus（VZV）in prokaryotic cells. Methods The recombinant plasmid pET28 a-gE-his containing gene encoding VZV-gE was constructed to transform into E.coli.The recombinant E.coli was cultured and induced by isopropyl-β-d-thiogalactoside（IPTG）and the gE protein was purified using affinity chromatography column and the specificity was examined by Western blot. Results The product of pET28 a-gE-his/BL21 induced by IPTG was separated from the mix proteins by the affinity chromatography column,the relative molecular mass of the expressed protein is about 60×10-3.Western blot confirmed the specificity of glycoprotein E. Conclusions The recombinant VZV-gE was successfully expressed in E.coli.Purification of VZV glycoprotein E with affinity chromatography is an effective method.
作者
高媛
吴克
梁锦
李津
肖杨
GAO Yuan;WU Ke;LIANG Jin;LI Jin;XIAO Yang(Bravovax Co.,Ltd,Wuhan,Hubei 430074,China)
出处
《中国病毒病杂志》
CAS
2018年第5期389-392,共4页
Chinese Journal of Viral Diseases
基金
武汉市科学技术计划项目应用基础研究计划(2016060101010038)
关键词
水痘-带状疱疹病毒
糖蛋白E
原核表达
亲和层析
Varicella-zoster virus
Glycoprotein E
Prokaryotic expression
Affinity chromatography
