猴B病毒囊膜蛋白gC基因密码子优化及其在大肠埃希菌中的表达

Optimization and Prokaryotic Expession of Gene Codon of Monkey B Virus Glycoprotein C

根据猴B病毒E2490标准株gC基因,选择大肠埃希菌偏好的密码子,设计合成了1 200个碱基,共编码400个氨基酸,测序证实后,克隆入pET-28b(+)载体,转化至大肠埃希菌BL21(DE3),在IPTG诱导下获得50 ku以包涵体形式表达的gC重组蛋白。Western blot分析表明表达产物能够被猴B病毒标准阳性血清识别。间接ELISA检测表明,重组蛋白与人单纯疱疹病毒1型、2型阳性血清无交叉反应。本研究为进一步建立检测猴B病毒抗体间接ELISA方法奠定了基础。

In accordance with the gene sequence of E2490 of Monkey B virus standard strains,the biased codon in E.coli was selected.The complete length of the synthesis gene was 1200bp,which encoded 400 amino acids.The constructed gene fragment was cloned into pET-28b（＋）,and transformed into E.coli BL21.The culture was induced by IPTG.The recombinant protein was mainly located in the inclusion body.The Western blot showed the protein could react specifically with the monkey B virus positive serum.The indirect ELISA showed no cross reactivity with sera of human herpes simplex Ⅰ,Ⅱ positive sera.The laid the foundation for make the establishment of indirect ELISA for the detection of monkey B virus and the development of DNA vaccine.