内源性血管生成抑制因子A rresten在E.coli JM109中的表达及抗新生血管生成的药理学研究

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,并进行表达,抑制新生血管的药理学实验中发现,该表达产物具有抑制鸡胚绒毛尿囊膜血管生长的功能。方法从健康产妇的胎盘组织中提取总RNA,经逆转录-聚合酶链式反应(RT-PCR)扩增出Arresten基因,构建重组质粒pBV220-Arr转化E.coli JM109进行原核表达,大量表达提取Arresten蛋白,用鸡胚绒毛尿囊膜实验进行活性测定。结果成功构建的重组质粒pBV220-Arr在E.coliJM109菌株中2~8 h均可获得表达,其中诱导4 h表达效率最高,Arresten蛋白可明显抑制鸡胚绒毛尿囊膜血管生长,活性功能明显强于血管抑素。结论成功构建Arresten基因重组质粒pBV220-Arr,并可在E.coli JM109菌株中获得表达,Arresten蛋白具有明显的抑制血管生成的作用。

Expression in E. coli JM109 and pharmacology on anti-angiogenesis of endogenetic angiogenesis inhibitor Arresten gene

Aim To construct prokaryotic expression vector for endogenetic angiogenesis inhibitor Arresten gene and to express recombinant.Pharmacology experiments found that the expression product had the function of restrain blood vessel growing on the chorioallantoic membrane(CAM).Methods The total RNA was extracted from the placenta organize of the normal puerpera,and Arresten gene was amplified by RT-PCR to construct recombinant vector pBV220-Arr.Then,it was transformed into E.coli JM109 for yielding recombinant human Arresten protein.The method to detect inhibiting angiogenesis activity was CAM study.Results The plasmid pBV220-Arr.could be expressed in E.coli JM109 from two hours to eight hours,and the expression yield reached the highest in two hours.The Arresten protein could restrain blood vessel growth of CAM evidently than that of angiostatin.Conclusion The recombinant plasmid pBV220-Arr was construted successfully and the target protein Arresten can express in E.coli JM109,the protein of Arresten had the evident effect of inhibite the activity of angiogenesis.