表达生存素相关microRNA-494基因腺病毒载体的构建

摘要

Objective To construct a recombinant adenovirus vector that expresses microRNA-494. Methods A correlation analysis between microRNA-494 and survivin mRNA was performed by Targetscan 5. 2. A fragment of gene was obtained by RT-PCR, which was connected to a linearized shuttle vector and cotransfected> after combined with genomic plasmid vector, into the 293 cells to produce the recombinant adenovirus vector for pre-microRNA-494 gene. The recombinant adenovirus vector was conformed and the protein expression was observed under fluorescent microscope. Results The recombinant adenovirus vector for pre-microRNA-494 gene was successfully constructed. Conclusion The recombinant adenovirus vector has been successfully constructed,which may be used in studying its effect on the expression of survivin mRNA in proastate cancer(PC-3) cells.%目的 构建表达生存索(survivin)相关rnicroRNA-494基因的重组腺病毒载体.方法 应用生物学软件Targetscan 5.2对microRNA-494基因与survivin mRNA作相关性分析.RT-PCR调取目的基因,将其连接线性化后的穿梭质粒载体,联合骨架质粒转染入293细胞后同源重组产生重组腺病毒载体.PCR及测序验证重组质粒,荧光显微镜下观察腺病毒荧光蛋白表达.结果 重组腺病毒载体构建成功.结论 成功构建了重组腺病毒载体,可用于研究其在前列腺癌PC-3细胞中对survivin基因表达的影响.