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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Discovery of virulence genes of Legionella pneumophila by using signature tagged mutagenesis in a guinea pig pneumonia model
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Discovery of virulence genes of Legionella pneumophila by using signature tagged mutagenesis in a guinea pig pneumonia model

机译：通过标记性诱变在豚鼠肺炎模型中发现嗜肺军团菌的致病基因

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摘要

Legionella pneumophila is the cause of Legionnaires' disease, which is a form of potentially fatal pneumonia. To identify genes required for virulence of the bacterium, a library of 1,386 L.pneumophila signature tagged transposon mutants was studied for guinea pig virulence. The mutants were screened in pools of 96 each in a guinea pig model of L.pneumophila pneumonia. Sixteen unique mutant clones were determined to have attenuated virulence after being screened twice in the animal model. All 16 mutants failed to multiply in both lungs and spleens. Four of the sixteen had no apparent defect for intracellular multiplication in macrophages. Partial DNA sequences of the interrupted genes adjacent to the transposon insertions showed that six of them had mutations in five known L.pneumophila virulence genes: dotB, dotF/icmG, dotO/icmB, icmX, and proA. Three of the sequenced clones contained mutations in genes without known homology to other published bacterial genes, and seven clones appeared to be homologous to five different known bacterial genes but are still being characterized. With this methodology, we demonstrate the existence of L.pneumophila genes responsible for non-macrophage-related virulence. The discovery of L.pneumophila virulence genes indicates the utility of the signature tagged mutagenesis technique for pulmonary pathogens.
机译：军团菌肺炎是军团病的病因，这是一种潜在的致命性肺炎。为了鉴定该细菌的毒性所需的基因，研究了1,386个嗜肺乳杆菌签名标签的转座子突变体库的豚鼠毒性。在肺炎支原体肺炎的豚鼠模型中，在每个96个池中筛选突变体。在动物模型中筛选两次后，确定了16个独特的突变克隆具有弱毒力。所有16个突变体均未能在肺和脾中繁殖。十六个中有四个在巨噬细胞中没有明显的细胞内增殖缺陷。与转座子插入物相邻的被打断基因的部分DNA序列显示，其中六个在五个已知的嗜肺乳杆菌毒力基因中具有突变：dotB，dotF / icmG，dotO / icmB，icmX和proA。其中三个已测序克隆中的基因突变与其他已发表的细菌基因没有已知同源性，七个克隆似乎与五个不同的已知细菌基因同源，但仍在表征中。用这种方法，我们证明了负责非巨噬细胞相关毒力的嗜肺乳杆菌基因的存在。 L.pneumophila毒力基因的发现表明签名标记的诱变技术对肺部病原体的实用性。

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