MicroRNA-9过表达载体的构建与鉴定

发布时间：2018-06-13 16:15

本文选题：microRNA + miR-　；参考：《现代生物医学进展》2015年26期

【摘要】：目的:MicroRNA(miRNA)是一类对基因表达起着转录后调控的小分子RNA,在正常发育和疾病发生过程中都发挥着重要的功能。其中,miR-9是一个序列高度保守的成员,在神经发育中调节神经干细胞的多种行为,包括分化、迁移等。但是,目前研究发现的miR-9的功能还存在一些相互矛盾和不清楚的地方,因此,我们拟构建miR-9的过表达载体,以便于对miR-9进一步的仔细研究。方法:我们采用分子克隆的常用方法,以pcDNA6.2-GW/miR为基础,构建出pcDNA6.2-GW/miR-9的表达质粒,分别用PCR、酶切和测序的方法验证质粒构建的正确性,并在Hela细胞和NIH3T3细胞中验证该质粒是否可以过表达miR-9小分子。结果:我们成功构建出正确的pc DNA6.2-GW/miR-9表达质粒,并且该质粒无论在人源的Hela细胞还是在小鼠源性的NIH3T3细胞中都可以过表达miR-9小分子。结论:构建了一个在不同种属间通用的miR-9过载体,为我们进一步研究miR-9的功能奠定了基础。
[Abstract]:Objective: microRNAs miRNAs are a class of small RNAs which play a role in normal development and disease development. MiR-9 is a highly conserved member that regulates neural stem cell behavior in neural development, including differentiation, migration, and so on. However, the functions of miR-9 found in the present study are still contradictory and unclear. Therefore, we intend to construct the over-expression vector of miR-9 for further careful study of miR-9. Methods: the expression plasmids of pcDNA6.2-GW / miR-9 were constructed based on pcDNA6.2-GW / miR-9 by molecular cloning methods, and the correctness of plasmid construction was verified by PCR, restriction endonuclease digestion and sequencing, respectively. The expression of miR-9 in Hela cells and NIH3T3 cells was examined. Results: we successfully constructed the correct expression plasmid of pc DNA6.2-GW / miR-9, which can overexpress miR-9 in both human Hela cells and mouse derived NIH3T3 cells. Conclusion: a general vector of miR-9 was constructed, which laid a foundation for further study on the function of miR-9.
【作者单位】：西安交通大学医学院附属红会医院;
【基金】：西安市红会医院院级科研基金项目(YJ2013013)
【分类号】：R3416;Q78

【共引文献】