【摘要】：豬小腸上皮細(xì)胞表面的產(chǎn)腸毒素大腸桿菌(ETEC)F4受體的基因型與其粘蛋白4(MUC4)基因密切相關(guān),根據(jù)MUC4基因內(nèi)含子7能否被限制性內(nèi)切酶XbaⅠ酶切,可以在基因水平上將所檢測(cè)的豬分為ETEC F4易感型純合子SS,易感型雜合子SR和抵抗型RR。為篩選對(duì)ETEC F4受體易感性和抗性仔豬用于本實(shí)驗(yàn)室的后續(xù)實(shí)驗(yàn),本實(shí)驗(yàn)利用MUC4基因的多態(tài)性,對(duì)送檢豬的樣品進(jìn)行初步的分型檢測(cè)。選取MUC4基因內(nèi)含子7不同等位基因型的仔豬,分離其小腸上皮細(xì)胞,并分別與野生型F4ab、F4ac、F4ad大腸桿菌,表達(dá)fae操縱子的重組大腸桿菌r F4ab、r F4ac、r F4ad進(jìn)行體外黏附和黏附抑制試驗(yàn)。研究結(jié)果表明,上述野生菌或重組菌對(duì)SS和SR兩種基因型的4周齡斷奶大白仔豬小腸上皮細(xì)胞均具有較好的黏附能力,而且經(jīng)過(guò)抗F4單克隆抗體處理后的細(xì)胞,對(duì)細(xì)菌的黏附數(shù)明顯下降。而RR型仔豬小腸上皮細(xì)胞不能黏附上述野生菌或重組菌。本研究為體外鑒定F4受體易感性仔豬,以及為進(jìn)一步研究ETEC F4的致病機(jī)理奠定了基礎(chǔ)和平臺(tái)。
[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.
【作者單位】： 揚(yáng)州大學(xué)獸醫(yī)學(xué)院;江蘇省動(dòng)物重要疫病與人獸共患病防控協(xié)同創(chuàng)新中心;揚(yáng)州大學(xué)動(dòng)物與科學(xué)學(xué)院;
【基金】：科技部畜禽重大疫病防控與高效安全養(yǎng)殖綜合技術(shù)研發(fā)重點(diǎn)專項(xiàng)(2016YFD0500900) 國(guó)家自然科學(xué)基金(31672579、30571374、30771603、31072136、31270171) 江蘇高校優(yōu)勢(shì)學(xué)科建設(shè)工程資助項(xiàng)目 中國(guó)農(nóng)業(yè)科學(xué)院哈爾濱獸醫(yī)研究所獸醫(yī)生物技術(shù)國(guó)家重點(diǎn)實(shí)驗(yàn)室開(kāi)放基金項(xiàng)目(SKLVBF201706)
【分類號(hào)】：S852.61

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