血管生成抑制因子arresten基因的克隆表达

Cloning and Expression of Human Arresten Gene

构建血管生成抑制因子arresten基因的原核表达重组体 ,并进行初步表达。从人胎盘组织中提取总RNA ,经反转录 聚合酶链式反应 (RT PCR)扩增出arresten基因 ;采用T A克隆法 ,将arresten基因克隆入pGEM T载体中 ,经DNA测序确认后 ,构建原核表达重组体pRSET Arr,转化大肠杆菌BL2 1 (DE3) ,用IPTG诱导表达。所获得的arresten基因经测序正确 ,并表达重组蛋白 ,经SDS PAGE分析 ,相对分子量为 2 6ku。构建的原核表达重组体pRSET Arr能高效表达重组arresten蛋白。

To construct prokaryotic expression recombinant of human arresten gene so as to express primarily recombinant arresten. Human arresten gene fragment was amplified from human placenta total RNA by RT PCR,and cloned into the vector of pGEM T by means of T A cloning. The sequence of arresten gene was analyzed. Prokaryotic expression vector pRSET Arr was constructed and transformed into E.coli DH5α. Recombinant arresten was expressed primarily in E.coli BL21(DE3) by IPTG induction. The sequence of human arresten gene obtained was correct. SDS PAGE analysis indicated that expressed product was about 26ku. The recombinant of pRSET Arr is an effective prokaryotic expression recombinant.\;