【摘要】 目的　在体外表达和纯化目的蛋白 ,为下一步抗攻击试验提供安全有效的产品。方法　将化学合成的恶性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BL2 1进行表达 :工程菌经超声破菌、离心、离子交换层析、疏水层析、分子筛层析等步骤纯化。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 2 3kDa ,占总菌体蛋白的 2 3 65 % ;纯度可达 95 %以上。Western -blot分析表明表达产物具有免疫原性。结论　成功构建pRSET -HGFSP重组表达系统并得到高效表达 ,纯化工艺令人满意更多还原

【Abstract】 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.更多还原