摘要
克隆和在大肠杆菌中表达人血管内皮生长因子１６５（ＶＥＧＦ１６５）。方法利用ＰＣＲ的方法从胎脑ｃＤＮＡ库扩增得到人ＶＥＧＦ１６５的全长ｃＤＮＡ并测定其核酸序列。利用基因重组技术构建ＶＥＧＦ１６５的原核表达载体，并在大肠杆菌中进行表达，用Ｗｅｓｔｅｒｎｂｌｏｔ检测表达产物。结果经测序表明，克隆的ＶＥＧＦ１６５与文献报道相符，Ｗｅｓｔｅｒｎｂｌｏｔ结果表明经用ＩＰＴＧ诱导后，ＶＥＧＦ１６５在大肠杆菌中表达出。结论克隆和表达出ＶＥＧＦ１６５。
Purpose To clone and express vascular endothelial growth factor 165(VEGF 165) in E.coli . Methods The cDNA of VEGF 165 was amplified with PCR from human fetus brain cDNA library.After sequenced,the VEGF 165 cDNA was cloned into procaryotic expression vector pTrc99A and the recombinant VEGF 165 expressed in E.coli JM 105, the expressed product was detected by western blot. Results The size of the product amplified by PCR was about 600 bp and the sequence was the same as the reported VEGF.The Western blot result showed that the VEGF165 protein was expressed in E.coli JM105. Conclusions VEGF165 was cloned and expressed.These results have laid the foundation for further studying of the function of VEGF 165.
出处
《上海医科大学学报》
CSCD
1999年第3期184-186,共3页
Journal of Fudan University(Medical Science)
基金
国家自然科学基金
上海生命中心资助
