中国畜牧兽医

• 生物技术 • 上一篇    下一篇

重组质粒pMEF2A在胎羊成纤维细胞中的表达

陈付英1,楚秋霞1,贾荣玲2,牛晖3,吴姣4,王二耀1,冯亚杰1,徐照学1   

  1. 1. 河南省农业科学院畜牧兽医研究所,河南郑州 450002;2. 河南省南阳农业职业学校,河南南阳 473000; 3.河南省肉牛工程技术中心有限公司,河南郑州 450003;4. 河南农业大学牧医工程学院,河南郑州 450003)
  • 收稿日期:2013-08-06 出版日期:2014-02-20 发布日期:2014-03-27
  • 通讯作者: 徐照学(1961—),教授。E-mail: xuzhaoxue11@163.com
  • 作者简介:陈付英(1973—),女,河南人,博士,助理研究员,研究方向:黄牛分子育种。 楚秋霞(1984—),女,河南人,硕士,助理研究员,研究方向:家畜胚胎工程。 陈付英、楚秋霞对本文具有同等贡献,并列为第一作者。
  • 基金资助:

    国家肉牛牦牛产业技术体系、肉牛健康养殖技术示范、优质转基因肉羊新品种培育(2014ZX08008003-003)。

Expression of the Recombinant Plasmid pMEF2A in Fetal Lamb Myoblasts

CHEN Fu-ying1, CHU Qiu-xia1, JIA Rong-ling2, NIU Hui3, WU Jiao4, WANG Er-yao1,FENG Ya-jie1, XU Zhao-xue1   

  1.  (1. Institute of Animal Husbandry and Veterinary Medicine, Henan Academy of Agricultural Sciences, Zhengzhou 450002,China;2. Nanyang Vocational School of Agriculture, Nanyang 473000,China;3. Research Center of Cattle Engineering Technology in Henan, Zhengzhou 450003,China;4. Institute of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450003,China)
  • Received:2013-08-06 Online:2014-02-20 Published:2014-03-27

摘要: 本研究用RT-PCR方法扩增并克隆了黄牛肌细胞增强因子2A(myocyte enhancer factor-2 A,MEF2A)基因CDS区,并将其插入到质粒载体pEGFP-C1的多克隆位点中,导入胎羊成纤维细胞中,同时转染空载体作对照,通过观测荧光蛋白的表达量检测转染效果。结果显示,转染48 h,空载体和携带目的基因的载体在胎羊成纤维细胞中均有表达,空载体的荧光表达量比目的基因的荧光表达量多。经过G418筛选,携带空载体的细胞能够获得稳定表达细胞系,携带目的基因细胞的筛选条件还需要做更深入的探索。

关键词: 肌细胞增强因子2A基因; 成纤维细胞; 真核表达

Abstract: The CDS of cattle myocyte enhancer factor-2 A (MEF2A) was amplified by RT-PCR and cloned into eukaryotic expression vector pEGFP-C1 and transfected into fetal lamb myoblasts, meanwhile transfection empty vector as control. The fluorescence intensity of transcription was detected by the reporter gene after transfection. The results indicated that the fluorescence intensity of cells with recombinant plasmid than that of cells with blank vector after 48 h transfection. By selection with G418, the stably transfected cell lines with blank vector were detected while cells with the screening condition of cells with recombinant plasmid need further optimize.

Key words: MEF2A gene; myoblasts; eukaryotic expression