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作 者:张岩 夏安婷 刘青悦 刘杰[1] 魏翔飞 曲波[1] 姜毓君[1] 王春梅[1] 张莉[1] ZHANG Yan;XIA Anting;LIU Qingyue;LIU Jie;WEI Qiangfei;QU Bo;JIANG Yujun;WANG Chunmei;ZHANG Li(The key laboratory of Dairy Science of Education Ministry,Northeast Agricultural University,Harbin 150030,China)
机构地区:[1]东北农业大学乳品科学教育部重点实验室,哈尔滨150030
出 处:《中国乳品工业》2021年第9期4-7,共4页China Dairy Industry
摘 要:以健康的泌乳期中国荷斯坦奶牛的乳腺上皮细胞(DCMECs)为实验模型,构建DNMT3A过表达载体pcDNA3.1-DNMT3A,用脂质体转染奶牛乳腺上皮细胞,采用荧光定量PCR技术检测奶牛乳腺上皮细胞中DNMT3A的mRNA表达,采用Bovine milk fat ELISA KIT检测细胞中的乳脂含量,采用Bovine lactose ELISA KIT检测细胞中的乳糖含量。结果表明,过表达DNMT3A后,奶牛乳腺上皮细胞生成的乳脂和乳糖含量均减少,表明DNA甲基化水平升高能够抑制奶牛乳腺上皮细胞乳脂和乳糖的合成与分泌。In this study,the mammary epithelial cells of healthy Chinese Holstein dairy cows during lactation were cultured and used as the experimental model.The DNMT3A overexpression vector pcDNA3.1-DNMT3A was constructed and then transfected into the dairy cow mammary epithelial cells.The mRNA expression of DNMT3A was detected by fluorescence quantitative PCR technology.The milk fat content in the cells was detected by Bovine milk fat ELISA KIT.The lactose content in the cells was detected by Bovine lactose ELISA KIT.The experimental results showed that after overexpression of DNMT3A,the milk fat content and the lactose content were reduced significantly in dairy cow mammary epithelial cells,indicating that the increasing of DNA methylation level can inhibit the synthesis and secretion of milk fat and lactose in dairy cow mammary epithelial cells。
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