狂犬病病毒G蛋白的原核表达及反应原性分析

为优化狂犬病病毒G蛋白的原核表达条件,探究其反应原性,参照狂犬病病毒CTN-1V10株编码G蛋白基因序列,采用生物工程合成的方法合成编码G蛋白的基因片段,并将该片段命名为G5F。将该片段与pET-28a原核表达载体连接,构建重组表达质粒pET-G5F,并将重组质粒进行诱导表达,在IPTG浓度为0.2 mmol/L、温度为20℃、诱导12 h时,重组蛋白表达量最高。SDS-PAGE电泳及Western blot分析表明,G5F重组蛋白可溶性表达量高,且可以被抗狂犬病病毒单克隆抗体识别,反应原性良好。

Prokaryoticexpression and Immunoreactivity Analysis of Rabies G Protein

To optimize the prokaryotic expression condition and the immunoreactivity of the rabies virus G protein(RABV G protein).Herein G protein coding genes section G5F was successfully synthesized with biological engineering synthesis method,using the rabies virus CTN-1V10 strain as a template.The recombinant plasmid was constructed by cloning the G5F gene into pET-28a,and then the plasmid was transformed into E.coli BL21 (DE3) competent cells.Highest protein expression level was achieved when induced with 0.2 mmol/L IPTG at 20 ℃ for 12 h.The results of SDS-PAGE and Western blot showed that RABV G protein was expressed as a soluble recombinant protein,and it could be recognized by antiRABV monoclonal antibody,which revealed a good immunoreactivity of this protein.