HBV PreS2-MBP融合蛋白在大肠杆菌中表达条件的优化

Optimization of Condition for Expression of HBV PreS2-MBP Fusion Protein in E. coli

目的优化HBV PreS2-MBP融合蛋白在大肠杆菌中的表达条件。方法通过改变重组质粒的宿主菌、培养基、诱导温度、诱导剂、诱导剂浓度、诱导时间等条件,利用SDS-PAGE和BandScan凝胶分析软件,分析以上条件改变对表达产物表达量的影响。将重组质粒连续传100代,检测融合蛋白的表达稳定性。结果重组质粒在大肠杆菌BL21菌株、GS培养基、28℃、0.5mmol/L IPTG或1.5mmol/L乳糖诱导4h时,目的蛋白的表达量最高,占菌体总蛋白的43.5%。重组质粒传代100代,融合蛋白的表达稳定。结论已确定出了融合蛋白在大肠杆菌中表达的最佳条件。

Objective To optimize the condition for expression of HBV PreS2-MBP fusion protein in E. coli. Methods Analyze the influence of host bacterium,culture medium,temperature and time for induction as well as inducer and its concentration on expression level of HBV PreS2-MBP fusion protein in E. coli by SDS-PAGE and BandScan software. Subculture recombinant E. coli strain for 100 passages and test the expression stability of HBV PreS2-MBP fusion protein. Results The expression level of HBV PreS2-MBP fusion protein reached 43.5% of total somatic protein in E. coli BL21 strain cultured in GS medium under induction of 0. 5 mmol/L IPTG or 1.5 mmol/L lactose at 28℃ for 4 h. The expression level of fusion protein in E. coli BL21 strain subcultured for 100 passages showed no significant difference with that in original strain. Conclusion The condition for expression of HBV PreS2-MBP fusion protein in E. coli was optimized.