猪miR-96-5p组织表达谱及其关键靶基因分析  

Tissue expression and analysis of swine miR-96-5p and its significant target genes

作　　者：杨敏[1] 滚双宝[1,2] 闫尊强 王鹏飞 杨巧丽[1] 黄晓宇 唐于然[1] 石海霞 马艳萍 赵进[4] YANG Min;GUN Shuangbao;YAN Zunqiang;WANG Pengfei;YANG Qiaoli;HUANG Xiaoyu;TANG Yuran;SHI Haixia;MA Yanping;ZHAO Jin(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Gansu Research Center for Swine Production Engineering and Technology,Lanzhou 730070,China;Library of Gansu Agricultural University,Lanzhou 730070,China;Animal Husbandry Development Service Station at Shizi Town in Lingtai County,Pingliang 744404,China)

机构地区：[1]甘肃农业大学动物科学技术学院,甘肃兰州730070 [2]甘肃省现代养猪技术工程研究中心,甘肃兰州730070 [3]甘肃农业大学图书馆,甘肃兰州730070 [4]甘肃省灵台县什字镇畜牧开发服务站,甘肃平凉744404

出　　处：《甘肃农业大学学报》2021年第5期18-27,共10页Journal of Gansu Agricultural University

基　　金：甘肃省青年科技基金计划项目(20JR5RA005);国家自然科学基金项目(31960646);甘肃省现代农业技术体系-猪鸡产业(GARS-ZJ-1);甘肃农业大学人才专项(GAU-KYQD-2018-27).

摘　　要：【目的】在前期感染C型产气荚膜梭菌(Clostridium perfringens type C,C.perfringens type C)攻毒组及对照组仔猪回肠组织高通量测序结果的基础上,为研究猪miR-96-5p在不同组织中的表达,分析其关键靶基因在仔猪感染C型产气荚膜梭菌导致腹泻过程中的调控作用.【方法】采用RT-qPCR方法检测miR-96-5p在仔猪各组织的表达量,构建组织表达谱,并检测对照组(IC)、易感组(IS)和耐受组(IR)回肠组织中miR-96-5p及其靶基因的表达量;利用生物信息学软件对miR-96-5p进行保守性分析、靶基因预测、GO功能和KEGG信号通路富集分析.【结果】miR-96-5p成熟体序列在不同物种间高度保守;预测到靶基因179个;预测获得与免疫炎症反应相关的信号通路有ErbB和MAPK;筛选到与仔猪抵抗C型产气荚膜梭菌感染相关的靶基因有FOXO1、MAP2K1;RT-qPCR结果显示,miR-96-5p在回肠组织中高表达,且其在IC、IS、IR组回肠组织中的表达量与高通量测序结果趋势一致,其靶基因FOXO1、MAP2K1在IR和IS组的表达量均极显著低于IC组(P<0.01),IS组极显著低于IR组(P<0.01).【结论】miR-96-5p可能靶向调控FOXO1、MAP2K1参与仔猪感染C型产气荚膜梭菌性腹泻的过程.【Objective】To investigate the tissue expression and analyze the key target genes of miR-96-5p,and to explore the regulatory mechanism of miR-96-5p in piglets diarrhea induced by Clostridium perfringens type C.【Method】The expression of miR-96-5p in piglet tissues was determined by using RT-qPCR to obtain tissue expression spectrum.The expression level of miR-96-5p in the ileum tissue was measured in the control group(IC),the susceptible group(IS)and the resistance group(IR).In addition,bioinformatics software was used to conduct conservative analysis,target gene prediction,GO terms and KEGG signal pathways enrichment of miR-96-5p.【Result】The mature sequence of miR-96-5p was highly conserved among different species and a total of 179 target genes were predicted.Signaling pathways related to immune inflammatory response ErbB and MAPK were obtained.Some target genes associated with resistance to Clostridium perfringens type C were selected,such as FOXO1 and MAP2K1.RT-qPCR results showed that miR-96-5p was highly expressed in ileum tissues,and its expression levels in the IC,IS and IR groups were consistent with the trend of high-throughput sequencing results.The expression level of target genes(FOXO1 and MAP2K1)in IR group and IS group was significantly lower than those in IC group(P<0.01),and that in IS group was significantly lower than in IR group(P<0.01).【Conclusion】The results suggest that miR-96-5p may target FOXO1 and MAP2K1 to participate in the regulation of diarrhea in piglets infected with Clostridium perfringens type C.

关 键 词：C型产气荚膜梭菌 miR-96-5p 仔猪 腹泻 

分 类 号：S828[农业科学—畜牧学]