人心室肌球蛋白轻链1基因的克隆及其体外的表达与纯化

Construction of pET22b human ventricular myosin light chain 1 and its expression in E. coli BL21

通过聚合酶链式反应方法扩增人心室肌球蛋白轻链1的基因片段,并将其克隆到pET-22b质粒,构建表达重组体,转化大肠杆菌BL21细胞,转化子细胞经IPTG诱导,目的蛋白在大肠杆菌中得到高效表达,凝胶成像扫描检测,其表达量接近细胞总蛋白的40%,经Ni2+亲和层析使目的蛋白得到纯化。

The cDNA of human ventricular myosin light chain 1 has been obtained by using the PCR method. Then it was inserted into pET22b to construct expression vector. High level expression of this target cDNA was observed in BL21 bacteria transformed with the recombinant vector after IPTG induction. The abundant VLC1 fusion protein constituted 40% of the total cellular protein. The fusion proteins were purified from bacterial lysate by using Ni^(2+ )affinity chromatography.