【摘要】 应用逆转录聚合酶链反应（RT－PCR法）从人脐静内皮细胞RNA中扩增了表皮生长因子（EGF）的cDNA，采用基因重组技术克隆到pGEM－3zf（＋）载体中，经全自动荧光测序仪测定了其序列。在PCR扩增引物上分别加有起始密码子（ATG）和终止密码子（TGA），以利于原核表达。经亚克隆，EGFcDNA克隆到表达载体pBV220的EcoRI、SmaⅠ位或上，在大肠杆菌DH5α中进行表达。SDS-聚丙烯酰胺凝胶电泳表明其表过量约占菌体总蛋白的10％。更多还原

【Abstract】 By means of reverse transcript polymerase chain reaction (RT-PCR) technology, a 171 bp nucleotide sequence that encodes human Epidermal growth factor (EGF) was obtained from cultured human vein endothial cells. The result sequencing EGF structural gene was identical with other known sequences. EGF cDNA was cloned in to DH5α on the plasmid pBV220 and a recombinant EGF was expressed successfully using PRPL promotor, accounting for 10% of total bacteri al protein, It’s molecular weight was about 6000, corresponding to the expected result.更多还原