重组人超氧化物歧化酶的基因克隆、表达及产物纯化研究

为了克隆人SOD-cDNA，构建表达载体，实现其在大肠杆菌中的高效稳定表达，通过抽提人肝组织总RNA，RT-PCR扩增人SOD cDNA，构建含rhSOD cDNA的表达质粒pLY-4/rhSOD，转化大肠杆菌JF1125进行表达研究。结果克隆到的rhSOD cDNA序列与文献报道一致，在宿主菌中获得高效表达，表达水平达68%以上；蛋白复性纯化工艺高效快速，rhSOD纯品纯度达98%以上，比活性达到2529u/mg;为用基因工程方法生产rhSOD打下基础。

Gene Cloning, Expression and Purification of Its Production of Recombinant Human Superoxide Dismutase

Human SOD cDNA was cloned and constructed an expression plasmid with high sufficient and stablility expresssion in E. coli. The rhSOD cDNA was amplified by RT-PCR with the template of the total RNA extracted from human liver tissue. The expression plasmid, pLY-4/rhSOD, containing rhSOD cDNA, was transformed into the E. coli JF1125. The sequence of the cloned rhSOD cDNA was identified with the reported data. The expression level reached to more than 68% of total bacteria proteins; The technology for protein renature and purification was efficiency and fast. The purity of the final products reached more than 98%. The value of bloactivity was determined as 2529 u/mg. This study gave enough support for production of rhSOD by biotechnology.