重组人尿激酶原基因的克隆及工程菌的构建与鉴定

Human Prourokinase cDNA Gene Cloning and Engineered Strain Construction and Characterization

利用RT PCR技术 ,从人脐带静脉上皮细胞中克隆重组人尿激酶原基因 ,用表达质粒pET2 9a构建了重组质粒 pET2 9a/ prouk ,并转化至大肠杆菌BL2 1(DE3)中 ,作为工程菌 .经IPTG诱导表达 ,在 4 6kDa处有一明显表达条带 ,表达量为约占菌体总蛋白的 2 0 % .

The human recombinant prourokinase gene was cloned from the epithelial cells of human umbilical vein by RT PCR. The pET29a/prouk plasmid was constructed and expressed in E.coli BL21(DE3) strain by IPTG induction. The expression product with a 46 kDa was identified by SDS PAGE, and the protein accounted for 20% of the bacterial total protein as inclusion body. The engineered strain showed the stability of its plasmid maintenance, resuscitation and expression efficiency in storage and regerenation.