畜牧·兽医

猪Gasp1和Gasp2基因对C2C12细胞分化的影响

  • 樊爽爽 ,
  • 张雪梅 ,
  • 刘晓贺 ,
  • 刘姣扬 ,
  • 明胜利 ,
  • 褚贝贝 ,
  • 杨国宇 ,
  • 王 江
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  • 河南农业大学 农业部动物生化与营养重点开放实验室,河南 郑州 450002
樊爽爽(1991-),女,河南荥阳人,在读硕士研究生,研究方向:动物生物化学。E-mail:990461231@qq.com

收稿日期: 2017-08-16

  网络出版日期: 2018-03-15

基金资助

农业部转基因重大专项(2014ZX0801015B);农业部948重点计划(2011-G35)

摘要

为研究猪Gasp1、Gasp2对成肌细胞分化的影响,采用RT-PCR技术从猪肌肉组织中扩增Gasp1和Gasp2基因片段,将其克隆入phiC31载体,构建重组质粒 phiC31-Gasp1和phiC31-Gasp2。将重组质粒转染至C2C12小鼠成肌细胞,经嘌呤霉素筛选在荧光显微镜下观察,获得稳定表达Gasp1和Gasp2基因的细胞系后,应用CCK-8法检测细胞增殖能力。分别在细胞分化1、2、3、4、5 d时收集细胞,采用实时荧光定量PCR检测MyoD、MyoG、P21及MHC肌细胞分化相关基因的表达变化,进而分析Gasp1和Gasp2基因对肌肉分化的影响。结果显示,本试验成功克隆了猪Gasp1和Gasp2基因,构建了phiC31- Gasp1和phiC31-Gasp2重组质粒,荧光显微镜下可见phiC31载体和Gasp1、Gasp2基因共转染的C2C12细胞表达强烈的绿色荧光,且对细胞的生长增殖无明显影响;实时荧光定量PCR检测结果表明,猪Gasp1和Gasp2可以促进细胞分化因子MyoD、MyoG、P21、MHC基因的表达。以上结果表明,Gasp1、Gasp2基因可以促进肌肉分化。

本文引用格式

樊爽爽 , 张雪梅 , 刘晓贺 , 刘姣扬 , 明胜利 , 褚贝贝 , 杨国宇 , 王 江 . 猪Gasp1和Gasp2基因对C2C12细胞分化的影响[J]. 河南农业科学, 2018 , 47(3) : 121 -127 . DOI: 10.15933/j.cnki.1004-3268.2018.03.024

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