《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (10): 2553-2559.doi: 10.16431/j.cnki.1671-7236.2016.10.007

• 生物技术 • 上一篇    下一篇

L型氨基酸转运载体1对奶牛乳腺中β-酪蛋白表达的影响

邢伟楠, 段晓宇, 杨洋, 侯晓明, 林叶   

  1. 东北农业大学生命科学学院, 哈尔滨 150030
  • 收稿日期:2016-01-26 出版日期:2016-10-20 发布日期:2016-10-28
  • 通讯作者: 林叶 E-mail:linlu516@163.com
  • 作者简介:邢伟楠(1990-),女,黑龙江哈尔滨人,硕士生,研究方向:泌乳生物学与乳腺功能调控,E-mail:1138815408@qq.com
  • 基金资助:

    国家自然科学基金(31401109);黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT-2015010);东北农业大学青年才俊基金资助项目(14QZ03)

Effect of L Type Amino Acid Transporter 1 on β-casein Expression in Mammary Epithelial Cells of Dairy Cows

XING Wei-nan, DUAN Xiao-yu, YANG Yang, HOU Xiao-ming, LIN Ye   

  1. College of Life Science, Northeast Agricultural University, Harbin 150030, China
  • Received:2016-01-26 Online:2016-10-20 Published:2016-10-28

摘要:

为研究L型氨基酸转运载体1(L type amino acid transporter 1,LAT1)对奶牛乳腺中β-酪蛋白表达的作用,本试验采用PCR技术体外扩增奶牛LAT1基因并构建LAT1真核表达载体,采用脂质体转染技术将LAT1真核表达载体转染奶牛乳腺上皮细胞,并于转染48 h后采用Western blotting技术检测LAT1过表达后乳腺上皮细胞中LAT1、4F2hc和β-酪蛋白的表达变化。荧光显微镜检测结果显示,LAT1真核表达载体成功转染奶牛乳腺上皮细胞;Western blotting检测结果显示,LAT1过表达的奶牛乳腺上皮细胞中LAT1极显著增加(P<0.01),β-酪蛋白的表达显著升高(P<0.05),4F2hc表达变化不显著(P>0.05)。这些结果提示LAT1对奶牛乳腺上皮细胞乳蛋白合成具有促进作用。

关键词: 奶牛; 乳腺上皮细胞; L型氨基酸转运载体1; β -酪蛋白

Abstract:

In order to investigate the effect of LAT1 on β-casein expression in mammary epithelial cells of dairy cows,the LAT1 gene of dairy cow was amplified by PCR and pIRES2-EGFP-LAT1 expression vector was constructed.Then pIRES2-EGFP-LAT1 expression vector was transfected into mammary epithelial cells.The effects of LAT1 overexpression on expression of LAT1,4F2hc and β-casein were detected by Western blotting after 48 h of transfection.The results showed that pIRES2-EGFP-LAT1 was transfected into mammary epithelial cells successfully.The expression of LAT1 was extremely significantly increased (P<0.01) and β-casein was significantly increased (P<0.05) when LAT1 was overexpressed,while the expression of 4F2hc was not significantly changed (P>0.05).These results suggested that LAT1 had an important role in promoting milk protein synthesis in mammary epithelial cells of dairy cow.

Key words: dairy cow; mammary epithelial cell; LAT1; β-casein

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