猪传染性胃肠炎病毒感染相关猪源miRNA的筛选及表达差异分析

Screening and differentials expression analysis of porcine transmissible gastroenteritis virus infection related of porcine miRNA

经生物信息学预测发现猪传染性胃肠炎病毒(TGEV)3′UTR可能是ssc-miR-182的潜在作用位点,本研究以猪传染性胃肠炎病毒为研究对象,构建TGEV 3′UTR双荧光素酶报告基因载体,并将其与合成的ssc-miR-182模拟体瞬时共转染至猪小肠上皮细胞,通过荧光监测仪测定荧光素酶活性;同时通过荧光定量PCR方法检测TGEV感染猪小肠上皮细胞后在不同时间段ssc-miR-182的表达变化情况。研究结果显示,ssc-miR-182对TGEV 3′UTR有一定的抑制作用,并且TGEV在感染小肠上皮细胞以后ssc-miR-182的表达明显上调。结果表明,宿主ssc-miR-182与TGEV的感染和复制增殖相关。

With porcine transmissible gastroenteritis virus as the research object,TGEV 3＇UTR dual luciferase reporter gene vectors were constructed.The bioinformatics prediction in TGEV 3＇UTR was the likeliest potential targets of miRNA,recombinant plasmids and the synthetic of miRNA and it＇s inhibitor in transient co-transfection of porcine intestinal epithelial cells,luciferase activity was measured by fluorescence monitor;moreover,research on the difference of miRNA expression on TGEV transmissible swine intestinal epithelial cells in different time periods.The result shown that the main site of action in microRNA CSFV the 3′-UTR,MicroRNA targets in ssc-miR-182 for TGEV 3′UTR has a certain extent,Meanwhile TGEV infection of swine intestinal epithelial cells after 24,36 and 48hexpression levels are rising,with healthy controls more than two-fold difference in multiples in 36 h,it is shown that ssc-miR-182 and TGEV infection has obvious relevance.