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荧光报告系统验证miR-142-5p的靶基因Becn1

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摘要 目的验证miR-142-5p的靶基因——自噬相关基因Becn1。方法生物信息学预测miR-142-5p与Becn1的结合位点,构建含有结合位点的Becn1基因3'UTR野生型和突变型质粒,将重组质粒与miR-142-5p-mimics或miR-NC共转染C2C12细胞,检测荧光素酶活性。结果 miR-142-5p与Becn1的3'UTR存在一个结合位点(123~129);成功构建野生型重组质粒p GL3-Becn1-3'UTR和靶位点含3个碱基突变的重组质粒Mut-3'UTR;与共转染p GL3-Becn1-3'UTR和miR-NC组相比,共转染p GL3-Becn1-3'UTR和miR-142-5p-mimics组荧光素酶活性显著下降,约为41.2%(P〈0.01);而与对照组相比,共转染Mut-3'UTR和miR-142-5p-mimics组的荧光素酶活性下降不明显(P〉0.05)。结论初步验证自噬相关基因Becn1是miR-142-5p的靶基因。
出处 《中国老年学杂志》 CAS CSCD 北大核心 2016年第6期1284-1287,共4页 Chinese Journal of Gerontology
基金 国家自然科学基金项目(No.81170327) 广东省医学科研基金(No.A2014470) 东莞市科技计划项目与医疗卫生一般项目(No.2012108102022,201410515200159) 广东医学院科研基金项目(No.XK1412,ZZDC006,ZZDC009,STIF201102)
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