猪TLR5基因表达水平与F18大肠杆菌侵染关系分析

doi:10.16431/j.cnki.1671-7236.2018.05.005

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (5): 1163-1169.doi: 10.16431/j.cnki.1671-7236.2018.05.005

猪TLR5基因表达水平与F18大肠杆菌侵染关系分析

赵呈祥¹, 戴开宇¹, 杜雨露¹, 包文斌^1,2, 吴圣龙^1,2

1. 扬州大学动物科学与技术学院, 江苏省动物遗传繁育与分子设计重点实验室, 扬州 225009;
2. 江苏省种猪繁育和健康养殖工程技术研究中心, 扬州 225009

收稿日期:2017-10-24 出版日期:2018-05-20 发布日期:2018-05-19
通讯作者: 吴圣龙 E-mail:slwu@yzu.edu.cn
作者简介:赵呈祥(1994-),男,江苏常州人,硕士生,研究方向:猪抗病育种,E-mail:862034465@qq.com
基金资助:
国家自然科学基金（31772560）；江苏省重点研发计划（现代农业）（BE2016305）；江苏省研究生科研创新计划项目（XKYCX17_062）

Analysis of the Relationship Between Expression Level of Porcine TLR5 Gene and Infection of Escherichia coli F18

ZHAO Chengxiang¹, DAI Kaiyu¹, DU Yulu¹, BAO Wenbin^1,2, WU Shenglong^1,2

1. Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
2. Jiangsu Engineering Research Center for the Reproduction and Healthy Breeding of Boar, Yangzhou 225009, China

Received:2017-10-24 Online:2018-05-20 Published:2018-05-19

摘要/Abstract

摘要：

为了探究猪Toll样受体（Toll-like receptor 5，TLR5）基因表达水平与F18大肠杆菌抗性的关系，试验通过不同血清型产肠毒素大肠杆菌（F18ab和F18ac）侵染猪小肠上皮细胞（IPEC-J2），同时通过脂多糖（LPS）分别诱导IPEC-J2细胞4和8 h，利用实时荧光定量PCR检测TLR5基因表达水平变化，并利用Western blotting进行蛋白表达分析。结果显示，不同血清型大肠杆菌（F18ab和F18ac）菌体侵染IPEC-J2细胞后，TLR5基因表达水平均极显著上调（P<0.01）；LPS诱导IPEC-J2细胞4和8 h后，TLR5基因表达水平均极显著上调（P<0.01），且在LPS诱导IPEC-J2细胞8 h后，TLR5基因表达水平明显高于诱导4 h。与对照组相比，细胞中TLR5蛋白的表达水平极显著上调（P<0.01），与LPS诱导及F18大肠杆菌菌体刺激IPEC-J2细胞后mRNA表达水平结果相一致。本研究在细胞水平上分析了TLR5表达水平和F18大肠杆菌侵染的相关性，进一步证实猪TLR5基因的表达水平在细胞抵抗F18大肠杆菌的侵染过程中发挥了重要的调控作用，为今后关于TLR5基因功能及其在大肠杆菌腹泻遗传育种应用的研究奠定基础。

关键词: 猪; TLR5基因; 大肠杆菌; mRNA表达; Western blotting

Abstract:

In order to explore the relationship between Toll-like receptor 5 (TLR5) gene expression and resistance to E.coli F18,the bacterial stimulation experiments were conducted in this study.Intestinal epithelial cells (IPEC-J2) were induced by lipopolysaccharide (LPS) and E.coli F18 with different serotypes (F18ab and F18ac),the mRNA expression level of TLR5 gene was detected by Real-time PCR,and the protein expression was detected by Western blotting.The results showed that the expression level of TLR5 gene was extremely significantly up-regulated (P<0.01) after IPEC-J2 cells were infected with different serotypes of E.coli (F18ab and F18ac),which was also extremely significantly up-regulated after IPEC-J2 cells were induced by LPS for 4 and 8 h (P<0.01).After IPEC-J2 cells were induced by LPS for 8 h,the expression level of TLR5 gene increased more significantly than induction for 4 h.After LPS and bacterial stimulation,the expression level of TLR5 protein in cells was extremely significantly up-regulated (P<0.01),which was consistent with the results of mRNA expression.This study further confirmed that the expression level of porcine TLR5 gene played an important role in the process of cell resistance to E.coli F18 infection by studying the relationship between the expression level of TLR5 gene and E.coli.It also laid the foundation for future research on TLR5 gene function and its application in genetic breeding of E.coli diarrhea.

Key words: porcine; TLR5 gene; E. coli; mRNA expression; Western blotting

中图分类号:

S828

赵呈祥, 戴开宇, 杜雨露, 包文斌, 吴圣龙. 猪TLR5基因表达水平与F18大肠杆菌侵染关系分析[J]. 《中国畜牧兽医》, 2018, 45(5): 1163-1169.

ZHAO Chengxiang, DAI Kaiyu, DU Yulu, BAO Wenbin, WU Shenglong. Analysis of the Relationship Between Expression Level of Porcine TLR5 Gene and Infection of Escherichia coli F18[J]. , 2018, 45(5): 1163-1169.

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猪TLR5基因表达水平与F18大肠杆菌侵染关系分析

Analysis of the Relationship Between Expression Level of Porcine TLR5 Gene and Infection of Escherichia coli F18

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