鸡卵清蛋白基因启动子克隆及其活性的检测

Cloning and activity analysis of the promoter of chicken ovalbumin gene

为了获得鸡输卵管特异表达的卵清蛋白基因启动子,研究在采用PCR方法从鸡心基因组DNA中扩增卵清蛋白基因启动子(5OV)的基础上,将其与p Ac GFP1-N1载体连接构建真核表达载体p Ac GFP1-5OV,并采用脂质体法将p Ac GFP1-5OV分别转染鸡输卵管上皮细胞和鸡成纤维细胞后,观察其在两种细胞中的表达活性。结果表明:利用克隆获得的5OV构建的真核表达载体p Ac GFP1-5OV在转染鸡输卵管上皮细胞后第24小时,在荧光倒置显微镜下可见绿色荧光,而转染鸡成纤维细胞后未观察到GFP表达。说明所克隆的5OV具有在输卵管上皮细胞特异表达的活性。

To obtain ovalbumin gene promoter of chicken oviduct - specific expression, ovalbumin gene promoter （5OV） was amplified from the DNA of genome in chicken heart using PCR assay. The amplified 5OV was connected with pAcGFP1 vector to construct a pAcGFPI - 5OV vec- tor. The pAcGFP1 -50V was used to transfect into chicken oviduct epithelial cells and fibroblasts using a liposome method, respectively. And then the expression activity of pAcGFP1 -5OV was observed in the two kinds of cells. The results showed that the green fiuorescence was ob- served under an inverted fluorescence microscope at 24 h after using the eukaryotic expression vector pAcGFP1 - 5OV to transfect chicken oviduct epithelial cells, but the expression of green fluorescent protein wasnt observed after transfeeting chicken fibroblasts. The results indicate that the cloned 5OV has specific expression activity in chicken oviduct epithelial cells.