幽门螺杆菌的全基因组转座子诱变及必需基因分析
Global transposon mutagenesis and essential gene analysis of Helicobacter pylori.
作者信息
Salama Nina R, Shepherd Benjamin, Falkow Stanley
机构信息
Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA.
出版信息
J Bacteriol. 2004 Dec;186(23):7926-35. doi: 10.1128/JB.186.23.7926-7935.2004.
Abstract
We have constructed a genome-saturating mutant library of the human gastric pathogen Helicobacter pylori. Microarray tracking of transposon mutants (MATT) allowed us to map the position of 5,363 transposon mutants in our library. While we generally found insertions well distributed throughout the genome, 344 genes had no detectable transposon insertions, and this list is predicted to be highly enriched for essential genes. Comparison to the essential gene set of other bacteria revealed a surprisingly limited overlap with all organisms tested (11%), while 55% were essential in some organisms but not others. We independently verified the essentiality of several gene products, including an HtrA family serine protease, a hypothetical protein with putative phospholipase D activity, and a riboflavin specific deaminase. A limited screen for motility mutants allowed us to estimate that 4.5% of the genome is dedicated to this virulence-associated phenotype.
摘要
我们构建了人类胃部病原体幽门螺杆菌的全基因组饱和突变体文库。转座子突变体微阵列追踪（MATT）使我们能够确定文库中5363个转座子突变体的位置。虽然我们通常发现插入在整个基因组中分布良好，但有344个基因未检测到转座子插入，预计该列表中富含必需基因。与其他细菌的必需基因集比较发现，与所有测试生物的重叠惊人地有限（11%），而55%在某些生物中是必需的，但在其他生物中不是。我们独立验证了几种基因产物的必需性，包括一个HtrA家族丝氨酸蛋白酶、一个具有假定磷脂酶D活性的假设蛋白和一个核黄素特异性脱氨酶。对运动性突变体的有限筛选使我们估计基因组的4.5%与这种与毒力相关的表型有关。
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