ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2018, Vol. 49 ›› Issue (5): 897-906.doi: 10.11843/j.issn.0366-6964.2018.05.004

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Relationship between Methylation in Promoter Region of TLR5 Gene and Resistance to Escherichia coli F18 of Sutai Weaned Piglets

DAI Chao-hui1, FENG Hai-yue1, ZONG Qiu-fang1, WU Sheng-long1,2, BAO Wen-bin1,2*   

  1. 1. 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:2017-10-23 Online:2018-05-23 Published:2018-05-23

Abstract:

The aim of this study was to investigate the regulation function of methylation in promoter region of TLR5 gene on the resistance to E. coli F18 of weaned piglets. The differential expressions of TLR5 gene and protein in small intestine (duodenum and jejunum) between E. coli F18-resistant and E. coli F18-sensitive Sutai weaned piglets were detected by qPCR and Western blot, respectively. The core promoter region, CpG islands and their acting elements of TLR5 were determined by bioinformatics analysis and dual luciferase reporter system, and the correlation between the methylation degree in promoter region and the expression of TLR5 gene in E. coli F18-resistant and E. coli F18-sensitive Sutai weaned piglets was detected and analyzed. The results showed that expression levels of TLR5 gene in duodenum and jejunum of sensitive individuals were significantly and very significantly higher than that in resistant individuals (P<0.05 and P<0.01), respectively. And the protein expressions of TLR5 in duodenum and jejunum of sensitive individuals were significantly higher than that of resistant individuals (P<0.05). The core promoter region of TLR5 gene included 2 CpG islands and 16 acting elements, the methylation level of the mC-6 site in the second CpG island of the promoter region had a certain regulation effect on the expression of TLR5 gene. The site located in the core promoter bound by transcription factor Sp1. The above results indicated that TLR5 gene played an important regulating role in the process of E. coli invasion, whose low expression was conducive to the piglets' resistance to E. coli. The methylation of the mC-6 CG site in the second CpG island of the TLR5 gene promoter could inhibit the binding of the transcription factor Sp1, which inhibited the expression of the TLR5 gene and ultimately affected the resistance of weaned piglets to E. coli.

Key words: Sutai pig, TLR5 gene, methylation, Escherichia coli F18, correlation analysis

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