摘要
通过PCR方法从pSEAP2-control质粒获得人胎盘碱性磷酸酯酶(PLAP)基因,并克隆到鸡输卵管特异性表达载体(pAB-Sig-SV40)卵清蛋白基因调控区下游。pSEAP2-control和pAB-Sig-AP-SV40质粒转染鸡胚细胞和乳腺癌细胞(MCF-7),转染48h后以对硝基苯磷酸钠(pNPP)为底物检测PLAP的活性,结果pSEAP2-control在鸡胚细胞和MCF-7细胞中表达PLAP,pAB-Sig-AP-SV40在鸡胚细胞中不表达,而在MCF-7细胞中表达。说明鸡胚细胞可以表达有活性的人源糖蛋白,另外也说明在雌激素受体细胞中,鸡卵清蛋白基因启动子可启动外源基因表达。
The coding sequence of human placental alkaline phosphatase(PLAP) gene was amplified from the pSEAP2-control plasmid by polymerase chain reaction(PCR). The plap gene was subcloned into chicken oviduct-specific expression vector(pAB-Sig-SV40). pSEAP2-control and pAB-Sig-AP-SV40 were transfected into chick embryo cells and MCF-7 cells. The results showed that expression of pSEAP2-control was achieved in both cells. That the recombinant vector was expressed in MCF-7 cells,but not in chicken embryo cells. These results indicated that human glycoprotein could be expressed in chicken embryo cells, and it had normal enzymatic activity. In addition,the chicken ovalbumin promoter could drive exogenous gene expression in the cells with estrogen receptor.
出处
《中国家禽》
北大核心
2007年第15期17-19,23,共4页
China Poultry
