pCold^(TM)TF载体原核表达FocA可溶性蛋白的研究

Prokaryotic Expression of Soluble FocA Protein with pCold ^(TM) TF Vector

为了较好地表达和纯化尿道致病性大肠杆菌F1C菌毛主要亚单位Foc A蛋白,研究以p Cold ^(TM) TF为载体,在大肠杆菌BL21(DE3)中原核表达F1C菌毛主要亚单位Foc A,验证表达的蛋白是否为可溶性蛋白,并对foc A基因表达的最佳条件进行了优化。结果表明:目的基因foc A在p Cold表达系统中于15℃下诱导24 h蛋白表达量最高,IPTG浓度在0.1~1.0 mmol/L范围内都能表达目的基因,且不同浓度对目的蛋白表达量无明显差异;表达出来的Foc A蛋白为可溶性蛋白。

In order to express and purify F1C fimbriae mainly subunit FocA protein from uropathogenic Escherichia coli, this experiment expressed FocA protein in Escherichia coli BL21 （DE3） with pColdTM TF vector, verifiedits solublility, and optimized focA gene expression condition. The results showed that focA protein expression reached the highest in pCold expression system by IPTG induced for 24 h under 15℃, and there were no significant difference in expression under 0.1-1.0 mmol/L IPTG conditions. Moreover it was found that the target protein FocA-TF was soluble.