FAK基因靶向miRNA表达质粒的构建及鉴定

目的构建靶向黏着酶（FAK）基因的miRNA真核表达质粒，为其在胃癌相关研究中的应用奠定基础。方法设计合成两条针对FAK基因的特异性miRNA干扰序列，定向克隆到pcDNATM6．2-GW／EmGFP—miR真核表达载体上，经鉴定后转染胃癌BGC-823细胞，实时定量PCR及Western—blot技术鉴定重组体对细胞FAK基因表达的干扰效果。结果针对FAK基因的两组miRNA干扰质粒构建成功，转染BGC-823细胞后，细胞FAKmRNA及FAK蛋白表达均明显降低。结论针对FAK基因的miRNA真核表达质粒成功构建，该质粒可用于miRNA进行靶向FAK的相关研究。

Construction and identification of miRNA expression vector targeting FAK gene

Objective To construct miRNA expression vector targeting FAK gene,and to lay the foundation for its application in the related studies on gastric carcinoma.Methods Two specific miRNA sequences targeting human FAK gene were designed and synthesized,and then were cloned into the pcDNATM6.2-GW/EmGFP-miR vector.The correct sequence was confirmed.The recombinant vector were transfected into BGC-823 cells and then the interference efficiency was observed by real-time PCR and western-blot test.Results pcDNA 6.2-GW...