Designer arrays for defined mutant analysis to detect genes essential for survival of Mycobacterium tuberculosis in mouse lungs
Designer arrays for defined mutant analysis to detect genes essential for survival of Mycobacterium tuberculosis in mouse lungs
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DOI：
10.1128/iai.73.4.2533-2540.2005
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发表时间：
2005-04-01
影响因子：
3.1
通讯作者：
Bishai, WR
中科院分区：
文献类型：
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作者：
Lamichhane, G;Tyagi, S;Bishai, WR
结核分枝杆菌诱发疾病的机制是复杂的，涉及体内生长和存活所需的大量细菌基因。为了鉴定这些基因，我们利用高通量微阵列检测方法快速筛选数百种独特的、基因型定义的转座子突变体，以获得高度特异性和灵敏度的体内存活。31米发现结核病基因是小鼠肺中体内存活所必需的。这些基因参与广泛的活动，包括代谢，细胞壁功能和调节。我们的筛选包括12个已知的分枝杆菌膜蛋白（mmpL）家族基因中的11个，这些基因中的6个-mmpL 4，mmpL 5，mmpL 7，mmpL 8，mmpL 10和mmpL 11-的突变严重损害了突变体在小鼠肺中繁殖的能力。这31个基因中的大多数在其他致病性分枝杆菌中是保守的，包括M。leprae和M.牛，这表明尽管分枝杆菌属的成员引起的人类病理学不同，但基本的体内存活机制的核心可能是高度保守的。在本文报道的31个基因中，有17个以前没有被描述过参与M.结核
The mechanisms by which Mycobacterium tuberculosis elicits disease are complex, involving a large repertoire of bacterial genes that are required for in vivo growth and survival. To identify such genes, we utilized a high-throughput microarray detection method to rapidly screen hundreds of unique, genotypically defined transposon mutants for in vivo survival with a high degree of specificity and sensitivity. Thirty-one M. tuberculosis genes were found to be required for in vivo survival in mouse lungs. These genes are involved in a broad range of activities, including metabolism, cell wall functions, and regulation. Our screen included 11 of the 12 known members of the mycobacterial membrane protein (mmpL) family genes, and mutation of 6 of these genes-mmpL4, mmpL5, mmpL7, mmpL8, mmpL10, and mmpL11-severely compromised the ability of the mutants to multiply in mouse lungs. Most of the 31 genes are conserved in other pathogenic mycobacteria, including M. leprae and M. bovis, suggesting that a core of basic in vivo survival mechanisms may be highly conserved despite the divergent human pathology caused by members of the mycobacterial genus. Of the 31 genes reported here, 17 have not been previously described to be involved in in vivo growth and survival of M. tuberculosis.