| 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 |
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| 引用本文： | 赵宇,罗艺晨,顾国靖,李博文,李文杰,周志雄,帅学宏,伍莉,陈吉轩,黄庆洲,焦寒伟.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[J].中国畜牧兽医,2019,46(10):2843-2850. |
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| 作者姓名： | 赵宇 罗艺晨 顾国靖 李博文 李文杰 周志雄 帅学宏 伍莉 陈吉轩 黄庆洲 焦寒伟 |
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| 作者单位： | 西南大学动物科学学院, 兽医科学工程研究中心, 荣昌 402460 |
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| 基金项目： | 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） |
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| 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Tnip1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Tnip1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Tnip1，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。
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| 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 |
| 收稿时间： | 2019-05-07 |
Target Gene Prediction and Functional Enrichment Analysis of mmu-miR-671-5p Mediated by Brucella |
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| ZHAO Yu,LUO Yichen,GU Guojing,LI Bowen,LI Wenjie,ZHOU Zhixiong,SHUAI Xuehong,WU Li,CHEN Jixuan,HUANG Qingzhou,JIAO Hanwei.Target Gene Prediction and Functional Enrichment Analysis of mmu-miR-671-5p Mediated by Brucella[J].China Animal Husbandry & Veterinary Medicine,2019,46(10):2843-2850. |
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| Authors: | ZHAO Yu LUO Yichen GU Guojing LI Bowen LI Wenjie ZHOU Zhixiong SHUAI Xuehong WU Li CHEN Jixuan HUANG Qingzhou JIAO Hanwei |
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| Institution: | College of Animal Sciences, Veterinary Scientific Engineering Research Center, Southwestern University, Rongchang 402460, China |
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| Abstract: | 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 Tnip1 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 Tnip1 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. |
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| Keywords: | Brucella RAW264 7 cells mmu-miR-671-5p target gene prediction functional enrichment analysis |
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