【摘要】 目的:探讨优化vMIP-II/MBP融合蛋白表达载体pMal在原核细胞中的表达条件,并对其表达产物进行纯化。方法:通过接菌、诱导表达,实验了解诱导时不同的温度、诱导时间、IPTG浓度对蛋白表达量的影响,并进行SDS-聚丙烯酰氨凝胶(SDS-PAGE)电泳分析。结果:发现该菌在含0 3mmol/LIPTG的TB培养基中,28℃诱导表达4h,可使诱导蛋白表达量占菌体蛋白总量的10%。结论:vMIP-Ⅱ/MBP的表达受温度条件影响较大,低温诱导时表达水平较高。从而为该工程菌的中试提供了实验基础。更多还原

【Abstract】 Aim: To optimize the expressing conditions of vMIP-II /MBP expressing vectors in primary cells and to purify its expressing pruducts. Methods: By planting bacteria?inducing expression,we study the influence of temperature?time and IPTG concentrations to expression amounts and do SDS-PAGE. Results: At the condition of 0 3 mmol/L IPTG? TB medium and inducing 4 h with 28℃, the expression amounts of fusion protein take account for 10% of the total protein. Conclusion: Temperature influence vMIP-Ⅱ/MBP fusion protein’s expression in transformed bacteria and the expression amounts are good at low temperature. This study provide the basis for the middle scale study.更多还原