抗菌肽hepcidin融合蛋白在大肠杆菌中表达条件的优化

Optimization of Expression Conditions of Antimicrobial Peptide Hepcidin Fusion Protein in E. coli

构建了his-hepcidin融合蛋白表达载体,转入大肠杆菌BL21(DE3)中,得到携带有pET-hec的E.coli BL2l(pET-Hec)菌株,为了确定融合蛋白在大肠杆菌BL21中的最佳表达条件,分别对诱导温度、诱导时机、诱导时间、诱导剂用量等进行了优化,通过Tricine-SDS-PAGE电泳分析,确定了最佳表达条件:37℃培养工程菌至OD600为0.6时,加入0.2mmol/LIPTG,25℃诱导表达8h。

In this study, the expression vector pET-Hec of fusion protein His-hepcidin was constructed and transferred into E. coli BL21 （DE3）and the E. coli BL21（pET-Hec） was obtained. In order to determine the best conditions for expression of the fusion protein, inducing temperature, opportunity, time, and dosage of inducer were tested. Through Trieine-SDS-PAGE eleetrophoresis analysis, the optimum expression conditions were determined as follow, adding 0.2 mmol/L IPTG while the OD600 reached 0.6 under 37℃ , and then inducing 8 hours under 25 ℃.