中国动脉硬化杂志2011，Vol.19Issue(5)：413-417,422,6.
肾素(前体)受体miRNA干扰质粒的构建及鉴定
Construction and Identification of miRNA Recombinant Eukaryotic Expression Vectors of (pro)Rennin Receptor
张娜娜 1李茂莲 1边云飞 1高奋 1杨慧宇 1肖传实1
作者信息
- 1. 山西医科大学第二医院,山西省太原市,030001
- 折叠
摘要
Abstract
Aim To construct and identify the miRNA eukaryotic expression vectors of (pro)renin receptor ( (P) RR). Methods miRNA nucletides of (P) RR were chemically synthesized and inserted into pcDNATM6 · 2-GW/EmGFPmiR vector, which were confirmed by sequencing, then the recombinant miRNA vectors were transfected into A7r5 cell by LipofectamineTM 2000. The mRNA expression of (P) RR was detected by Real-time PCR and Western blot.Results Sequencing suggested that miRNA eukaryotic expression vectors targeting (P) RR possessed correct read frame and nucleotide sequence, and green fluorescene of the transient transfected A7r5 cell lines could be observed under inverted fluorescence microscope. Real-time PCR and Western blot results showed that the sequence of X2-2-3 and X2-3-3 could effectively knockdown the level of mRNA and protein of (P) RR. Conclusions miRNA eukaryotic expression vectors targeting (P)RR were successfully constructed and the effectively interference RNA were identified, which may be used for understanding the effect of (P)RR in the vascular smooth muscle cells.关键词
肾素(前体)受体/微小干扰核糖核酸/血管平滑肌细胞Key words
(pro)Rennin Receptor/ miRNA/ Vascular Smooth Muscle Cell分类
生物科学引用本文复制引用
张娜娜,李茂莲,边云飞,高奋,杨慧宇,肖传实..肾素(前体)受体miRNA干扰质粒的构建及鉴定[J].中国动脉硬化杂志,2011,19(5):413-417,422,6.
