Identification of Yersinia enterocolitica genes affecting survival in an animal host using signature-tagged transposon mutagenesis
Identification of Yersinia enterocolitica genes affecting survival in an animal host using signature-tagged transposon mutagenesis
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DOI：
10.1046/j.1365-2958.1999.01324.x
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发表时间：
1999-04-01
影响因子：
3.6
通讯作者：
Miller, VL
中科院分区：
文献类型：
--
作者：
Darwin, AJ;Miller, VL
致病性耶尔森菌与哺乳动物宿主的局部和全身感染有关。在这项研究中，使用标记转座子诱变技术来鉴定感染小鼠模型中生存所需的小肠结肠炎耶尔森菌基因。筛选了大约2000个转座子插入突变体进行衰减。根据它们在混合感染中与野生型菌株竞争的能力判断，这导致鉴定出55个在动物宿主中生存缺陷的突变体。共有28个突变体在毒力质粒中插入转座子，验证了筛选结果。两个具有严重毒力缺陷的质粒突变体在一个未表征的区域插入。一些染色体插入是在一个基因簇参与o抗原的生物合成。其他染色体插入鉴定的基因先前未被证明是小肠结肠炎菌体内存活所必需的。这些基因包括参与外膜成分合成、应激反应和营养获取的基因。一个严重减毒的突变体插入了大肠杆菌的pspC基因（噬菌体休克蛋白C）的同源物。噬菌体休克蛋白操纵子在大肠杆菌中没有已知的生化或生理功能，但显然对大肠杆菌感染期间的存活至关重要。
Pathogenic Yersinia species are associated with both localized and systemic infections in mammalian hosts. In this study, signature-tagged transposon mutagenesis was used to identify Yersinia enterocolitica genes required for survival in a mouse model of infection. Approximately 2000 transposon insertion mutants were screened for attenuation. This led to the identification of 55 mutants defective for survival in the animal host, as judged by their ability to compete with the wild-type strain in mixed infections. A total of 28 mutants had transposon insertions in the virulence plasmid, validating the screen. Two of the plasmid mutants with severe virulence defects had insertions in an uncharacterized region. Several of the chromosomal insertions were in a gene cluster involved in O-antigen biosynthesis. Other chromosomal insertions identified genes not previously demonstrated as being required for in vivo survival of Y.enterocolitica. These include genes involved in the synthesis of outer membrane components, stress response and nutrient acquisition. One severely attenuated mutant had an insertion in a homologue of the pspC gene (phage shock protein C) of Escherichia coli. The phage shock protein operon has no known biochemical or physiological function in E. coli, but is apparently essential for the survival of Y.enterocolitica during infection.