重组人附睾特异蛋白BDFx的优化表达与纯化

Opitimized Expression and Purification of Recombinant Human Epididymis-Specific Gene BDFx Expressed in E.coli

通过改变异丙醇-β-D硫代半乳糖苷诱导浓度、诱导温度及诱导培养时间,优化含重组人附睾特异蛋白BDFx工程菌的表达条件.运用DEAE阴离子与CM阳离子交换层析及S-100分子筛层析纯化重组蛋白,用基质辅助激光解吸离子化-飞行时间质谱法、SDS-聚丙烯酰胺凝胶电泳、紫外扫描等方法鉴定目的蛋白.结果显示,缩短诱导培养时间、降低IPTG浓度等可提高重组蛋白的表达量,表明适当提高诱导表达温度不会形成包涵体,并可缩短表达时间.

Different concentrations of IPTG, inducing times and inducing temperatures are used to optimize the expression condition for recombinant BDFx in E. coli. The recombinant protein is purified by DEAE anion exchange-chromatography, CM positive ion exchange-chromatography, and molecular sieve chromatography, and the protein is identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, SDS- PAGE, and ultraviolet scanning. The results indicate that, shortening the cultural time and reducing the concentration of IPTG might be able to increase the expression amount of the recombinant protein. Increasing the inducing expression temperature properly can shorten the expression time and will not cause the forming of inclusion body.