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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
Objective To express and purify the target protein in vitro and provide a safe and effective product for the next anti-attack test. Methods The chemically synthesized Plasmodium falciparum protective antigen complex gene (HGFSP) was recombined with the expression vector pRSET and expressed in Escherichia coli BL21. The engineered bacteria were characterized by sonication, centrifugation, ion exchange chromatography, hydrophobic chromatography and molecular sieve layer Analysis and other steps of purification. Results SDS-PAGE showed that the expressed product was non-fusion and soluble. The relative molecular mass was 23kDa, accounting for 23.35% of the total bacterial proteins. The purity of the product was over 95%. Western-blot analysis showed that the expressed product was immunogenic. Conclusion The pRSET-HGFSP recombinant expression system was successfully constructed and expressed efficiently, and the purification process was satisfactory