【摘要】 从原代培养的人脐静脉血管内皮细胞 (HUVEC)提取细胞总RNA ,采用逆转录PCR (RT PCR)方法得到VEGF受体Flt 1胞外区前 3个IgG样区域cDNA片段 (Flt 1n3 )。将获得的受体基因克隆到真核表达载体 pcD NA3.1中 ,得到重组质粒 pcDNA3.1/Flt 1n3 ,通过脂质体转染方法将其转入中国仓鼠卵巢细胞 (CHO) ,用G418筛选得到稳定表达目的蛋白的细胞克隆。经固相结合实验筛选得到表达与VEGF特异结合的目的蛋白的细胞克隆 ,细胞测活结果表明 ,表达产物具有抑制人脐静脉内皮细胞 (HUVEC)增殖的活性。鸡胚测活结果表明 ,CHO细胞表达的rFlt 1n3 能抑制鸡胚绒毛尿囊膜的血管形成。更多还原

【Abstract】 The N terminal first three IgG like domain of VEGF receptor Flt 1 (flt l n3 ) was cloned from primary culture of human umbilical vascular endothelial cells (HUVEC) by RT PCR method. Flt l n3 was functionally expressed in stably transfected CHO cells by using recombinant plasmid PCF3, which was constructed by inserting Flt l n3 cDNA into eukaryotic expression vector pcDNA3.1. The product secreted in the medium can specifically bind to rh VEGF 165 and inhibit rh VEGF 165 stimulated HUVEC proliferation. Our results also showed the expressed Flt l n3 can inhibit angiogenesis of chick embryo chorio allantoic membrane(CAM).更多还原