猪miR-369靶基因肿瘤坏死抑制因子-α(TNF-α)的鉴定  

Identification of Porcine Tumor Necrosis Factor-α (TNF-α) as the Target Gene of the miR-369

作　　者：母治平[1] 邓娟[1,2] 王滔[1] 李明洲[1] 李学伟[1] 

机构地区：[1]四川农业大学动物科技学院动物遗传育种研究所,雅安625014 [2]重庆市潼南县畜牧局,潼南402660

出　　处：《农业生物技术学报》2012年第8期922-927,共6页Journal of Agricultural Biotechnology

基　　金：国家转基因生物新品种培育重大专项(No.2009ZX08009155B和No.2008ZX08006003)

摘　　要：microRNAs(miRNAs)是一类长约22nt的非编码小RNA分子,作为关键的转录后调控因子调控真核生物的基因表达,广泛参与细胞的生长发育、分化和凋亡等生物学过程。本研究通过生物信息学预测发现,猪(Susscrofa)肿瘤坏死抑制因子-α(TNF-α)的3'非编码区具有2个潜在的miR-369的靶位点。并进一步通过双荧光素酶报告系统分析,将猪TNF-α3'非编码区构建到双荧光素酶载体中,转染至猪髋动脉内皮细胞系,荧光素酶活性测定显示,miR-369过表达组的荧光比值显著低于对照组(P<0.05),而miR-369抑制组相比对照组没有显著差异(P=0.752)。研究结果提示,猪miR-369能靶向调控TNF-α的表达,对深入研究脂肪沉积性状重要候选基因TNF-α的转录后调控机制具有重要价值。microRNAs （miRNAs）, a class of small -22-nucleotide-long noncoding RNAs, emerge as key post-transcriptional regulators of gene expression in eukaryotic organisms and involve in a variety of cellular processes, such as cell proliferation, cell differentiation and apoptosis. In silico analysis of the tumor necrosis factor-α（TNF-α） and miR-369 showed that the 3＇UTR region of porcine （Sus scrofa） TNF-α adenylate/uridylate-rich elements （AREs） contained two putative miR-369 target sites. The porcine TNF-α 3＇UTR region was constructed into the dual-luciferase reporter vector, transfected in the porcine iliac artery endothelial cell （PIEC cell） line to measure the luciferase activity by dual-luciferase reporter assay. The results indicated that the luciferase activity of miR-369 mimics group was significantly lower than that of the control group （P〈0.05）. Nonetheless, there was not a significant differences between the inhibitors and the control groups （p=-0.752）. Our results demonstrated that the TNF-a should be the target gene ofmiR-369 and provides evidence for further research of post-transcriptional mechanisms of TNF--α which affect fat deposition.

关 键 词：猪 miR-369 肿瘤坏死抑制因子-α(TNF-α) 双荧光素酶报告基因系统 

分 类 号：Q255[生物学—细胞生物学]