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Analysis of firnbrial gene clusters and their expression in enterohaernorrhagic Escherichia coli O157 : H7

机译:肠出血性大肠杆菌O157:H7的肺基因簇及其表达分析

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The sequence of two enterohaemorrhagic Escherichia coli (EHEC) 0157:H7 strains reveals the possession of at least 16 fimbrial gene clusters, many of the chaperone/usher class. The first part of this study examined the distribution of these clusters in a selection of EHEC/EPEC (enteropathogenic E. coli) serotypes to determine if any were likely to be unique to E. coli 0157:H7. Six of the clusters, as determined by the presence of amplified main subunit or usher gene sequences, were detected only in the E. coli 0157 and 0145 serotypes tested. With the exception of one serotype 0103 strain that contained an Ipf2 cluster, Ipf sequences were only detected in E. coli 0157 of the serotypes tested. Expression from each cluster was measured by the construction of chromosomally integrated IacZ promoter fusions and plasmid-based eGFP fusions in E. coli 0157:H7. This analysis demonstrated that the majority (11/15) of main fimbrial subunit genes were not expressed under the majority of conditions tested in vitro. One of the clusters showing promoter activity, loc8, has a temperature expression optimum indicating a possible role outside the host. From the presence of pseudogenes in three of the clusters, the lack of FimH-like minor adhesins in the clusters and their limited expression in vitro, it would appear that E coli 0157:H7 has a limited repertoire of expressed functional fimbriae. This restricted selection of fimbriae may be important-in bringing about the tropism E coli 0157:H7 demonstrates for the terminal rectum of cattle.
机译:两个肠出血性大肠杆菌(EHEC)0157:H7菌株的序列显示拥有至少16个纤维基因簇,其中许多是伴侣/诱捕类。这项研究的第一部分检查了EHEC / EPEC(致病性大肠杆菌)血清型中这些簇的分布,以确定是否有可能是大肠杆菌0157:H7所特有的。通过扩增的主要亚基或引诱基因序列的存在确定的六个簇仅在测试的大肠杆菌0157和0145血清型中检测到。除了一个包含Ipf2簇的血清型0103菌株外,仅在测试的血清型的大肠杆菌0157中检测到Ipf序列。通过构建染色体整合的IacZ启动子融合体和大肠杆菌0157:H7中基于质粒的eGFP融合体来测量每个簇的表达。该分析表明,大多数(11/15)主要纤维亚基基因在体外测试的大多数条件下均未表达。显示启动子活性的簇之一,loc8,具有最佳温度表达,表明在宿主外可能发挥作用。从三个簇中假基因的存在,簇中缺乏FimH样次要粘附素及其在体外的有限表达来看,似乎大肠杆菌0157:H7具有有限的表达功能性菌群。对菌毛的这种有限选择可能很重要,因为它会导致大肠杆菌0157:H7向牛的终末直肠表现出嗜性。

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