畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (4): 508-512.

• 动物营养 • 上一篇    下一篇

stat5高效表达对奶牛乳腺上皮细胞泌乳能力的影响

刘晓飞,高学军*,李庆章,佟慧丽   

  1. 东北农业大学 乳品科学教育部重点实验室,哈尔滨 150030
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-04-20 发布日期:2011-04-20
  • 通讯作者: 高学军

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

LIU Xiao-fei, GAO Xue-jun*, LI Qing-zhang, TONG Hui-li   

  1. Key Laboratory of Dairy Science Ministry of Education, Northeast Agricultural University,Harbin 150030, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-04-20 Published:2011-04-20

摘要: 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pcDNA3.1+- 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Western Blotting 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。

关键词: stat5, 高效表达, 稳定转染, 奶牛, 乳腺上皮细胞, 泌乳

Abstract: 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 (P<0.01);β-casein, lactose and pSTAT5 protein were increased (P<0.05). The results showed that, eukaryotic expression vector pcDNA3.1+- stat5-αS1 could be highly expressed in DCMECs, and could raise lactation ability of DCMECs.

Key words: stat5, high expression, stable transfection, dairy cow, mammary gland epithelial cells, lactation