人miR-17-92基因簇载体构建及其在293T细胞中的表达验证

发布时间：2018-01-22 00:52

  本文关键词： miR--基因簇 T细胞 表达　出处：《生物技术通讯》2016年06期 　论文类型：期刊论文

【摘要】：目的:研究人miR-17-92基因簇在肾癌等疾病发生过程中的功能。方法:采用基因重组技术,将miR-17-92基因簇亚克隆至pc DNA4真核细胞表达载体,构建miR-17-92基因簇真核表达载体;通过脂质体转染的方法将miR-17-92的过表达质粒和对照质粒分别转染293T细胞,并用实时定量PCR技术验证转染24和48 h后细胞内pri-miR-17-92和miR-17、miR-18a、miR-19a、miR-19b1、miR-20a、miR-92a1等分子的相对过表达比例。结果:miR-17-92基因簇真核表达载体构建成功,并在293T细胞中实现了显著性过表达效果。结论:为进一步研究人miR-17-92基因簇在肾脏发育过程中的功能及其与肾脏癌症发病相关性的研究提供了实验基础。
[Abstract]:Objective: to study the function of human miR-17-92 gene cluster in renal cell carcinoma and other diseases. Methods: gene recombination technique was used. The miR-17-92 gene cluster was subcloned into the eukaryotic expression vector of PC DNA4, and the eukaryotic expression vector of miR-17-92 gene cluster was constructed. The overexpression plasmids of miR-17-92 and control plasmids were transfected into 293T cells by liposome transfection. The intracellular pri-miR-17-92 and miR-17 miR-18a miR-19a were confirmed by real-time quantitative PCR after transfection for 24 and 48 hours. The relative overexpression ratio of miR-19b1mmiR-20ahmiR-92a1 and other molecules was compared. Results the eukaryotic expression vector of the gene cluster of miR-17-92 was successfully constructed. Significant overexpression was achieved in 293T cells. Conclusion:. It provides an experimental basis for the further study of the function of human miR-17-92 gene cluster in renal development and its association with renal cancer.
【作者单位】： 山西大学生命科学学院;
【基金】：山西省高等学校科技创新项目(113548901027)
【分类号】：R737.11
【正文快照】： 现代社会,人们由于工作压力大、运动量少、长期高蛋白饮食等原因,使肾功能受到一定的影响,出现肾血管阻力下降、肾功血管舒张、肾小球滤过率明显增加等症状,严重情况下甚至诱发癌症的发生。在我国,肾癌是泌尿系统最常见的恶性肿瘤之一,其发病率仅次于膀胱癌,居泌尿系肿瘤第二，

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