中文摘要：

目的优化重组HIV-1跨膜蛋白Gp41包涵体的纯化条件。方法将表达Gp41的大肠杆菌菌体高压匀浆破碎、洗涤分离提取包涵体,以不同pH值的低浓度变性剂溶解包涵体,稀释复性并用离子交换层析纯化目的蛋白,纯化产物经West-ernblot进行鉴定。结果以50mmol/LTris buffer、2mol/L,pH11.5尿素溶解的包涵体蛋白得到很好的溶解,目的蛋白复性得率大于40%。经纯化后,目的蛋白的纯度大于90%,且具有良好的反应原性。结论优化了重组HIV-1跨膜蛋白Gp41包涵体的纯化条件,为HIV-1Gp41抗原纯化工艺的放大提供了依据。

英文摘要：

Objective To optimize the purification condition for inclusion body of recombinant HIV-1 transmembrane protein Gp41. Methods The recombinant E. coli in which Gp41 was expressed was disrupted by high-pressure homogenization and washed,and the inclusion body was isolated,dissolved with low concentration denaturing agent at various pH values,then refolded and purified by ion exchange chromatography. The purified target protein was identified by Western blot. Results More than 40% of inclusion body was refolded after dissolution with 50 mmol / L Tris buffer containing 2 mol / L urea at pH 11. 5. The target protein reached a purity of more than 90% after purification and showed good reactogenicity. Conclusion The purification condition for inclusion body of recombinant HIV-1 transmembrane protein Gp41 was optimized,which provided a basis for the scale-up of purification procedure of HIV-1 Gp41 antigen.