Construction and Identification of miRNA Recombinant Eukaryotic Expression Vectors of (pro)Rennin Receptor
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    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.

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ZHANG Na-Na, LI Mao-Lian, BIAN Yun-Fei, GAO Fen, YANG Hui-Yu, and XIAO Chuan-Shi. Construction and Identification of miRNA Recombinant Eukaryotic Expression Vectors of (pro)Rennin Receptor[J]. Editorial Office of Chinese Journal of Arteriosclerosis,2011,19(5):413-417.

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  • Received:January 21,2011
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