stat5高效表达对奶牛乳腺上皮细胞泌乳能力的影响

Effects on Lactation Ability of Dairy Cow Mammary Gland Epithelial Cells by stat5 High Expression

本研究旨在检验构建的stat5表达载体对奶牛乳腺上皮细胞泌乳能力的影响。构建真核表达载体pcD-NA3.1+-stat5-αS1,稳定转染到奶牛乳腺上皮细胞中。利用细胞活力分析仪、HPLC、Real-time PCR和WesternBlotting技术检测乳腺上皮细胞转染前后stat5基因和蛋白的表达量、细胞活力及乳糖和酪蛋白分泌情况。结果表明,与空白细胞和空白载体组相比,非磷酸化STAT5蛋白和stat5基因的表达量增加(P<0.01),乳糖含量提高(P<0.05),细胞活力和增殖能力增强(P<0.01),酪蛋白和磷酸化STAT5(pSTAT5)表达增多(P<0.05)。结果提示,构建的载体pcDNA3.1+-stat5-αS1在乳腺上皮细胞中高效表达,显著提高乳腺上皮细胞的泌乳能力。

The expression vector Stat5 was analysed for the change of effects on lactation ability of dairy cow mammary gland epithelial cells（DCMECs）.The expression vector was stably transfected by G418.The proliferations and viabilities of DCMECs,expression of β-casein,lactose,stat5 gene and STAT5 protein were detected by cell counter analyser,HPLC,Western Blotting and Real-time PCR.Compared with the control group,the proliferations and viabilities of DCMECs,expression of stat5 gene and STAT5 protein were both increased（P0.01）;β-casein,lactose and pSTAT5 protein were increased（P0.05）.The results showed that,eukaryotic expression vector pcDNA3.1＋-stat5-αS1 could be highly expressed in DCMECs,and could raise lactation ability of DCMECs.