不同启动子对于牛催乳素表达的调控作用

Regulation on the Expression of Bovine Prolactin Gene by Different Promoters

摘要:

在细胞水平上比较不同启动子对于牛催乳素(bPRL)表达的调控作用。分别构建了以CMV启动子、牛催乳素基因启动子和山羊b-酪蛋白基因启动子作为调控元件的bPRL真核细胞表达载体, 分别命名为pCMV、pPRLP和pP1A3。将3种载体分别转染小鼠垂体瘤细胞和小鼠乳腺上皮细胞, 使用RT-PCR和定量RT-PCR分析3种启动子启动bPRL在2种细胞系中的表达效果。pCMV在2种细胞中有效表达bPRL; pPRLP在2种细胞中的表达效果与pCMV接近; pP1A3不在垂体细胞中表达, 在乳腺细胞中表达。pP1A3具有乳腺表达特异性; pPRLP能够在垂体和乳腺中高表达, 在其他组织的表达特异性有待进一步研究。

Abstract:

To compare the regulation effects by different promoters on bovine prolactin gene expression in different cell lines, three recombinant bovine prolactin expression vectors were constructed using different promoters, i.e., CMV promoter, bovine prolactin gene promoter and goat β-casein gene promoter, respectively named pCMV, pPRLP and pP1A3, which were transfected into two cell lines, mouse pituitary tumor cell strain (AtT20) and mouse mammary epithelial cell strain (HC11), respectively. RT-PCR and real-time RT-PCR were used to investigate the expression level of the above three vectors in both cell lines. pCMV vector was effectively expressed in both cell lines, pPRLP vector had a similar expression level to that of pCMV in both cell lines, pP1A3 was expressed in HC11 but not in AtT20. pP1A3 was tissue-specific to mammary gland. pPRLP was able to express with a significant level in pituitary and mammary glands, while its tissue-specific characteristics in other tissues need further investigation.