摘要
为了探明miR-125a在精子发生中的功能,以小鼠基因组DNA为模板扩增miR-125a的前体片段(pre-miR-125a),经过双酶切后连接入质粒pcDNA3.1(+),构建miR-125a超表达载体(pcDNA3.1(+)-miR-125a);将pcDNA3.1(+)-miR-125a转染至GC-1spg细胞系,通过qRT-PCR和Western Blot验证其超表达效果。结果表明:PCR扩增得到约290bp的miR-125a前体片段,成功构建了pcDNA3.1(+)-miR-125a过表达载体,在GC-1spg生殖细胞系中约有3.5倍的超表达效果,为深入研究miRNAs在精子发生过程的功能及其机制奠定基础。
To elucidate the role of miR-125a during spermatogenesis in mice,miR-125a precursor fragments（pre-miR-125a）was amplified by PCR,and cloned into pcDNA3.1（＋）plasmid.pcDNA3.1（＋）-miR-125a was induced into the GC-1 spg cells to verify the effect of overexpression.The results showed as follows：the eukaryotic plasmid pcDNA3.1（＋）-miR-125a was successfully constructed.The expression of miR-125a in GC-1 spg cells induced with plasmid pcDNA3.1（＋）-miR-125a was increased by 3.5 times.In conclusion,a mouse miR-125a overexpression vector was constructed which is necessary for understanding its effects during spermatogenesis.
作者
宋峰峰
张恺阳
安俊辉
覃金洲
田秀娥
曾文先
SONG Fengfeng;ZHANG Kaiyang;AN Junhui;QIN Jinzhou;Tian Xiue;ZENG Wenxian(College of Animal Science and Technology,Northwest A ＆ F University, Yangling, Shaanxi 712100, China)
出处
《家畜生态学报》
北大核心
2018年第4期11-15,共5页
Journal of Domestic Animal Ecology
基金
国家自然科学基金项目(31072029
31272439)
