中文摘要：

目的：构建miR-139-5p低表达慢病毒载体,检测其在H9c2细胞的表达效果。方法根据大鼠miR-139-5p序列,设计合成其靶点序列,并合成寡核苷酸双链,克隆入慢病毒载体pGC-LV,与pHelper 1．0和pHelper 2．0共转染293T细胞,包装病毒。收取病毒上清液,纯化后感染 H9 c2细胞。

英文摘要：

Aim To construct a lentiviral low expres-sion of miR-139-5 p vector and validate its expression efficiency in H9c2 cells. Method Target sequence was designed according to the sequence of rat miR-139-5p. Oligonucleotide duplex was synthesized and cloned into the lentiviral vector pGC-LV. The recombi-nant lentiviral vector, pHelper 1. 0, and pHelper 2. 0 were co-transfected into 293T cells, packaging virus. Then H9 c2 cells were infected with the supernatant containing lentiviral particles, and its infection effi-ciency and miR-139-5 p expression were determined by fluorescent microscope and real-time quantitative PCR, respectively. Results A lentiviral low expression of miR-139-5p vector was successfully constructed. The infection efficiency in H9c2 cells reached over 95%, and the relative expression of miR-139-5p was significantly down-regulated. Conclusion The lentiviral low expression of miR-139-5p vector is successfully constructed, and the expression of miR-139-5p in infected H9c2 cells is inhibited effectively.