microrna125a质粒表达载体构建及其表达活性.pdf

　　中国组织工程研究 第 16 卷 第2 期 2012 –01 –08 出版

　　Chinese Journal of Tissue Engineering Research January 8, 2012 Vol.16, No.2

　　MicroRNA-125a质粒表达载体的构建及其表达活性★

　　张 涛，李 东，李 华，高 辉

　　Construction and expressing activity of microRNA-125a plasmid expression vectors

　　Zhang Tao, Li Dong, Li Hua, Gao Hui

　　Abstract

　　BACKGROUND: pSuper is a kind of eukaryotic expression vectors of non-coding RNA. It expresses the precursor of miRNA in Center of Tumor

　　plasma, and then the precursor is cut by Dicer enzyme to produce mature miRNA. This process completely imitates the formation Diagnosis and

　　process of endogenous miRNA. In this study, pSuper is utilized to construct an expression vector of miR-125a, which could inhibit Treatment, Chengdu

　　Millitary General

　　liver cancer. It provides basis for the further investigation on antitumor mechanism. Hospital of Chinese

　　OBJECTIVE: To construct the eukaryotic plasmid pSuper-miR-125a expressing miR-125a and to confirm the expressing PLA, Chengdu

　　efficiency and specificity of miR-125a in the plasmid. 610083, Sichuan

　　Province, China

　　METHODS: Target fragments in the precursor of miR-125a Pre-miR-125a were amplified by PCR. The fragments were cloned

　　into pMD18-T transition plasmid using T -A cloning method. The target fragments were cut by restriction enzyme and then Zhang Tao★, Master,

　　connected to pSuper plasmid to construct recombinant plasmid pSuper-miR-125a. The recombinant plasmid was transfected into Chief physician,

　　Hela cells. The expressing efficiency and specificity of miR-125a in the recombinant plasmid was detected by real-time PCR. Center of Tumor