内源性血管生成抑制因子Arresten的序列测定及在E.coli BL21中的表达

Sequencing of Arresten,an endogenous angiogenesis inhibitor gene,and its expression in E.coli BL21

目的构建内源性血管生成抑制因子Arresten基因的原核表达载体,并对其进行序列测定和在E.coli BL21中进行表达。方法从健康产妇的胎盘组织中提取总RNA,经反转录-聚合酶链式反应(RT-PCR)扩增出Arresten基因,对其序列测定后构建重组质粒pBV220-Arr转化E.coli BL21进行原核表达。结果成功筛选出Arresten基因,并将基因序列和蛋白序列提交基因库,成功构建了重组质粒pBV220-Arr,重组质粒在菌株中获得表达。结论构建的原核表达重组体pBV220-Arr能高效表达重组Arresten蛋白。

Objective To sequence the endogenous angiogenesis inhibitor Arresten gene and to express its recombinant in E. coli BL21 with a constructed prokaryotic expression vector. Methods Human Arresten gene was amplified by RT-PCR using total RNAs extracted from placenta tissue of healthy puerpera. The recombinant plasmid pBV220-Arr was sequenced and transfected into E.coli BL21 after being constructed and expressed by this prokaryotic-expression system. Results The Arresten gene was screened successfully and submitted to the GenBank. Recombinant plasmid pBV220-Arr was constructed and expressed in the BL21 successfully. Conclusion The pBV220-Arr was demonstrated highly efficient for expression of Arresten proteins.