布鲁氏菌介导的mmu-miR-671-5p的靶基因预测与功能富集性分析  

Target Gene Prediction and Functional Enrichment Analysis of mmu-miR-671-5p Mediated by Brucella

作　　者：赵宇[1] 罗艺晨[1] 顾国靖 李博文[1] 李文杰 周志雄 帅学宏 伍莉[1] 陈吉轩 黄庆洲[1] 焦寒伟 ZHAO Yu;LUO Yichen;GU Guojing;LI Bowen;LI Wenjie;ZHOU Zhixiong;SHUAI Xuehong;WU Li;CHEN Jixuan;HUANG Qingzhou;JIAO Hanwei(College of Animal Sciences,Veterinary Scientific Engineering Research Center,Southwestern University,Rongchang 402460,China)

机构地区：[1]西南大学动物科学学院兽医科学工程研究中心

出　　处：《中国畜牧兽医》2019年第10期2843-2850,共8页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金青年基金项目(31802215);重庆市自然科学基金项目(cstc2018jcyjA0807);中央高校基本科研业务费项目(XDJK2019C024、XDJK2019D013)

摘　　要：为进行布鲁氏菌介导的mmu-miR-671-5p的靶基因预测,本试验利用布鲁氏菌(Brucella)感染RAW264.7细胞后,分别运用miRanda和TargetScan软件进行差异表达的mmu-miR-671-5p靶基因的预测,预测的靶基因分别有11 953和9 252个,将预测结果取交集进行韦恩(Venn)分析,重叠部分的靶基因有3 681个;利用GO与KEGG进行功能富集性分析,再利用PicTar软件进一步对mmu-miR-671-5p的靶基因进行预测,将预测结果与Venn分析结果进一步取交集,结果显示,mmu-miR-671-5p的预测靶基因有7个;实时荧光定量PCR分别验证7个预测靶基因的相对表达量发现,Tnfrsf1b与Tnip 1基因相对表达量极显著降低(P<0.01);在RAW264.7细胞中转染mmu-miR-671-5p inhibitors,分别验证7个预测靶基因的相对表达量发现,Tnfrsf1b与Tnip 1基因的相对表达量极显著升高(P<0.01);对mmu-miR-671-5p与Tnfrsf1b和Tnip1的3′非翻译区(3′UTR)结合靶位点分别进行预测,结果初步表明,mmu-miR-671-5p的靶基因为Tnfrsf1b与Tnip 1,其预测结合靶位点均只有1个,分别位于Tnfrsf1b和Tnip1 3′UTR全长位置的91和305 bp。本研究结果为进一步揭示mmu-miR-671-5p在布鲁氏菌感染RAW264.7细胞过程中的功能提供科学依据。In order to predict the target gene of mmu-miR-671-5p mediated by Brucella,the differentially expressed mmu-miRNA-671-5p target genes were predicted by miRanda and TargetScan softwares after RAW264.7 cells were infected by Brucella.The predicted target genes were 11 953 and 9 252,respectively.The predicted results of the two softwares were intersected and analyzed by Venn which showed 3 681 overlapping target genes.The functional enrichment were analyzed by GO and KEGG,and then PicTar software was used to further predict the target gene of mmu-miRNA-671-5p.The predicted results were further intersected with the results of Venn which showed that there were 7 target genes was the finally predicted results of mmu-miRNA-671-5p.The relative expression of 7 predictive target genes was verified by Real-time quantitative PCR.The results showed that the relative expression of Tnfrsf1b and Tnip 1 genes were extremely significantly decreased(P<0.01).And then the mmu-miRNA-671-5p inhibitors were transfected into RAW264.7 cells and the 7 predictive targets were verified.The results showed that the relative expression of Tnfrsf1b and Tnip 1 genes was extremely significantly increased(P<0.01),and the target sites of mmu-miRNA-671-5p and the 3′UTR of Tnfrsf1b and Tnip1 were predicted respectively.The results preliminarily showed that the target genes of mmu-miRNA-671-5p were Tnfrsf1b and Tnip1.Both of them had only one predictive binding site,located at 91 and 305 bp of the full-length positions of 3′UTR of Tnfrsf1b and Tnip1,respectively.This study provided a scientific basis for further revealing the function of mmu-miRNA-671-5p in Brucella infected RAW264.7 cells.

关 键 词：布鲁氏菌 RAW264.7细胞 mmu-miR-671-5p 靶基因预测 功能富集性分析 

分 类 号：Q78[生物学—分子生物学]