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

doi:10.16431/j.cnki.1671-7236.2019.10.004

中国畜牧兽医 ›› 2019, Vol. 46 ›› Issue (10): 2843-2850.doi: 10.16431/j.cnki.1671-7236.2019.10.004

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

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

西南大学动物科学学院, 兽医科学工程研究中心, 荣昌 402460

收稿日期:2019-05-07 出版日期:2019-10-20 发布日期:2019-10-21
通讯作者: 焦寒伟 E-mail:jiaohanwei@swu.edu.cn
作者简介:赵宇(1996-),男,贵州贵阳人,硕士生,研究方向:人兽共患传染病学与动物功能基因组学,E-mail:812912001@qq.com;罗艺晨(1981-),女,重庆人,硕士,实验师,研究方向:中兽医学,E-mail:93442859@qq.com赵宇与罗艺晨对本文具有同等贡献,并列为第一作者
基金资助:
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）

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

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

Received:2019-05-07 Online:2019-10-20 Published:2019-10-21

摘要/Abstract

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。

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

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.

Key words: Brucella; RAW264.7 cells; mmu-miR-671-5p; target gene prediction; functional enrichment analysis

中图分类号:

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.

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 and Veterinary Medicine, 2019, 46(10): 2843-2850.

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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

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