| 张正平 赵勤鹏 黄琳红 王栋琪 许正伟 郭华 昌震 刘团江.MicroRNA-9 过表达载体的构建与鉴定[J].,2015,15(26):5033-5037 |
| MicroRNA-9 过表达载体的构建与鉴定 |
| Construction and Identification of microRNA-9 Overexpression Vector |
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| DOI： |
| 中文关键词: microRNA miR-9 过表达载体 pcDNA6.2-GW/miR 载体 |
| 英文关键词: microRNA miR-9 Overexpression plasmid pcDNA6.2-GW/miR plasmid |
| 基金项目:西安市红会医院院级科研基金项目（YJ2013013） |
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| 中文摘要: |
| 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pcDNA6.2-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。 |
| 英文摘要: |
| Objective:MicroRNA (miRNA) is a kind of small RNA that post-transcriptionally regulates gene expression and plays
an important role in normal development and disease development. miR-9 is a highly conserved miRNA and regulates many behaviors of
neural stem cells, including differentiation and migration etc. Some discrepancies and ambiguities, however, still exist about the function
of miR-9. Therefore, we aimed to construct the overexpression vector of miR-9 to facilitate the detailed research of miR-9.Methods:We
first employed molecular clone methods to construct the pcDNA6.2-GW/miR-9 expression plasmid based on a linear plasmid named
pcDNA6.2-GW/miR. Then the plasmid was tested by PCR, restriction digestion and sequencing. Last, the expression of miR-9 by
pcDNA6.2-GW/miR-9 plasmid was verified by transfecting it into Hela cells and NIH3T3 cells.Results:We contructed right pcDNA6.
2-GW/miR-9 plasmid and that pcDNA6.2-GW/miR-9 plasmid could overexpress miR-9 both in human cell line Hela and in murine cell
line NIH3T3.Conclusion:We successfully constructed a miR-9 overexpression vector, which could be used to overexpress miR-9 in
several cells of different origins. This research provided a basis for our further detailed analysis of the function of miR-9. |
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