恶性疟原虫多表位重组疫苗在大肠杆菌中的表达及纯化

目的 在体外表达和纯化目的的蛋白，为下一步抗攻击试验提供安全有效的产品。方法 将化学合成的恶生疟原虫保护性抗原复合基因（ＨＧＦＳＰ）与表达载体ｐＲＳＥＴ重组，在大肠杆菌ＧＩ２１进行表达；工程菌经超声破菌、离心、离子交换层析、疏水层析、分子支析等步骤纯化。结果 ＳＤＳ－ＰＡＧＥ显示表达产物以非融合、可溶性的形式表达，相对分子质量为２３ｋＤａ，占总菌体蛋白的２３．６５％，纯度可达９５％以上。Ｗｅｓｔｅ

Expression and purification of hybrid genes of protective antigens of plasmodium falciparum in non-fusion system

Objective To express and purify the target protein in vitro and to obtain safer and more efficient product for further anti-challenge test. Methods The chemically synthesized hybrid genes of protective antigens(HGFSP) was recombinated with pRSET expression vector and expressed in E coli BL 21 . The E coli was lysed by ultrasonication. Supernatant of the lysate was purified by ion-exchange chromatography, reversed chromatography and gel filtration. Results SDS-PAGE analysis suggested the target protein was expressed in soluble and non-fusion form after induced by IPTG. Its molecular weight was 23kDa and consisted of 23.65% of the total bacterial proteins. Purity of the target protein was above 95% after purification. Western blotting showed that the target protein had immunogenicity. Conclusion The pRSET-HGFSP expression recombinant has been successfully constructed and can be highly expressed in E coli BL 21 . The result of purification was satisfying.