F4大肠杆菌黏附素受体易感性仔猪的体外鉴定研究
发布时间：2018-08-04 10:32
【摘要】：猪小肠上皮细胞表面的产肠毒素大肠杆菌(ETEC)F4受体的基因型与其粘蛋白4(MUC4)基因密切相关,根据MUC4基因内含子7能否被限制性内切酶XbaⅠ酶切,可以在基因水平上将所检测的猪分为ETEC F4易感型纯合子SS,易感型杂合子SR和抵抗型RR。为筛选对ETEC F4受体易感性和抗性仔猪用于本实验室的后续实验,本实验利用MUC4基因的多态性,对送检猪的样品进行初步的分型检测。选取MUC4基因内含子7不同等位基因型的仔猪,分离其小肠上皮细胞,并分别与野生型F4ab、F4ac、F4ad大肠杆菌,表达fae操纵子的重组大肠杆菌r F4ab、r F4ac、r F4ad进行体外黏附和黏附抑制试验。研究结果表明,上述野生菌或重组菌对SS和SR两种基因型的4周龄断奶大白仔猪小肠上皮细胞均具有较好的黏附能力,而且经过抗F4单克隆抗体处理后的细胞,对细菌的黏附数明显下降。而RR型仔猪小肠上皮细胞不能黏附上述野生菌或重组菌。本研究为体外鉴定F4受体易感性仔猪,以及为进一步研究ETEC F4的致病机理奠定了基础和平台。
[Abstract]:The genotype of enterotoxigenic Escherichia coli (ETEC) F4 receptor on the surface of porcine intestinal epithelial cells is closely related to its mucin 4 (MUC4) gene. According to whether intron 7 of MUC4 gene can be digested by restriction enzyme Xba I, The tested pigs can be classified into susceptible homozygotes SSF4, susceptible heterozygotes SR and resistant RRs at the gene level. In order to screen ETEC F4 receptor susceptible and resistant piglets for further experiment in our laboratory, the polymorphism of MUC4 gene was used to detect the genotyping of pig samples. Small intestinal epithelial cells were isolated from piglets with different allelic genotypes of intron 7 of MUC4 gene. The recombinant E. coli r F4abtr F4actr F4ad expressing fae operon was used for in vitro adhesion and adhesion inhibition test. The results showed that the two genotypes of SS and SR had a good adhesion ability to the intestinal epithelial cells of weaned white piglets, and the cells were treated with anti-F4 monoclonal antibody. Adhesion to bacteria decreased significantly. The intestinal epithelial cells of RR piglets could not adhere to the above wild bacteria or recombinant bacteria. This study laid a foundation and platform for identification of susceptibility to F4 receptor in vitro and further study on pathogenesis of ETEC F4 in piglets.
【作者单位】： 扬州大学兽医学院;江苏省动物重要疫病与人兽共患病防控协同创新中心;扬州大学动物与科学学院;
【基金】：科技部畜禽重大疫病防控与高效安全养殖综合技术研发重点专项(2016YFD0500900) 国家自然科学基金(31672579、30571374、30771603、31072136、31270171) 江苏高校优势学科建设工程资助项目 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室开放基金项目(SKLVBF201706)
【分类号】：S852.61
本文编号：2163652
[Abstract]:The genotype of enterotoxigenic Escherichia coli (ETEC) F4 receptor on the surface of porcine intestinal epithelial cells is closely related to its mucin 4 (MUC4) gene. According to whether intron 7 of MUC4 gene can be digested by restriction enzyme Xba I, The tested pigs can be classified into susceptible homozygotes SSF4, susceptible heterozygotes SR and resistant RRs at the gene level. In order to screen ETEC F4 receptor susceptible and resistant piglets for further experiment in our laboratory, the polymorphism of MUC4 gene was used to detect the genotyping of pig samples. Small intestinal epithelial cells were isolated from piglets with different allelic genotypes of intron 7 of MUC4 gene. The recombinant E. coli r F4abtr F4actr F4ad expressing fae operon was used for in vitro adhesion and adhesion inhibition test. The results showed that the two genotypes of SS and SR had a good adhesion ability to the intestinal epithelial cells of weaned white piglets, and the cells were treated with anti-F4 monoclonal antibody. Adhesion to bacteria decreased significantly. The intestinal epithelial cells of RR piglets could not adhere to the above wild bacteria or recombinant bacteria. This study laid a foundation and platform for identification of susceptibility to F4 receptor in vitro and further study on pathogenesis of ETEC F4 in piglets.
【作者单位】： 扬州大学兽医学院;江苏省动物重要疫病与人兽共患病防控协同创新中心;扬州大学动物与科学学院;
【基金】：科技部畜禽重大疫病防控与高效安全养殖综合技术研发重点专项(2016YFD0500900) 国家自然科学基金(31672579、30571374、30771603、31072136、31270171) 江苏高校优势学科建设工程资助项目 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室开放基金项目(SKLVBF201706)
【分类号】：S852.61
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