Global transposon mutagenesis and essential gene analysis of Helicobacter pylori

被引:242
作者
Salama, NR
Shepherd, B
Falkow, S
机构
[1] Fred Hutchinson Canc Res Ctr, Div Human Biol, Seattle, WA 98109 USA
[2] Stanford Univ, Sch Med, Dept Microbiol & Immunol, Stanford, CA 94305 USA
关键词
D O I
10.1128/JB.186.23.7926-7935.2004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:7926 / 7935
页数:10
相关论文
共 54 条
[31]  
Ji YD, 2002, METHOD ENZYMOL, V358, P123
[32]   Identification and characterization of Helicobacter pylori genes essential for gastric colonization [J].
Kavermann, H ;
Burns, BP ;
Angermüller, K ;
Odenbreit, S ;
Fischer, W ;
Melchers, K ;
Haas, R .
JOURNAL OF EXPERIMENTAL MEDICINE, 2003, 197 (07) :813-822
[33]   Large-scale identification of essential Salmonella genes by trapping lethal insertions [J].
Knuth, K ;
Niesalla, H ;
Hueck, CJ ;
Fuchs, TM .
MOLECULAR MICROBIOLOGY, 2004, 51 (06) :1729-1744
[34]   Essential Bacillus subtilis genes [J].
Kobayashi, K ;
Ehrlich, SD ;
Albertini, A ;
Amati, G ;
Andersen, KK ;
Arnaud, M ;
Asai, K ;
Ashikaga, S ;
Aymerich, S ;
Bessieres, P ;
Boland, F ;
Brignell, SC ;
Bron, S ;
Bunai, K ;
Chapuis, J ;
Christiansen, LC ;
Danchin, A ;
Débarbouillé, M ;
Dervyn, E ;
Deuerling, E ;
Devine, K ;
Devine, SK ;
Dreesen, O ;
Errington, J ;
Fillinger, S ;
Foster, SJ ;
Fujita, Y ;
Galizzi, A ;
Gardan, R ;
Eschevins, C ;
Fukushima, T ;
Haga, K ;
Harwood, CR ;
Hecker, M ;
Hosoya, D ;
Hullo, MF ;
Kakeshita, H ;
Karamata, D ;
Kasahara, Y ;
Kawamura, F ;
Koga, K ;
Koski, P ;
Kuwana, R ;
Imamura, D ;
Ishimaru, M ;
Ishikawa, S ;
Ishio, I ;
Le Coq, D ;
Masson, A ;
Mauël, C .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (08) :4678-4683
[35]  
Manoil C, 2000, METHOD ENZYMOL, V326, P35
[36]   Living dangerously:: How Helicobacter pylori survives in the human stomach [J].
Montecucco, C ;
Rappuoli, R .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2001, 2 (06) :457-466
[37]  
MUSHEGIAN AR, 1996, P NATL ACAD SCI USA, V93, P1026
[38]   Urease plays an important role in the chemotactic motility of Helicobacter pylori in a viscous environment [J].
Nakamura, H ;
Yoshiyama, H ;
Takeuchi, H ;
Mizote, T ;
Okita, K ;
Nakazawa, T .
INFECTION AND IMMUNITY, 1998, 66 (10) :4832-4837
[39]   Cloning and genetic characterization of Helicobacter pylori catalase and construction of a catalase-deficient mutant strain [J].
Odenbreit, S ;
Wieland, B ;
Haas, R .
JOURNAL OF BACTERIOLOGY, 1996, 178 (23) :6960-6967
[40]   Optimized BlaM-transposon shuttle mutagenesis of Helicobacter pylori allows the identification of novel genetic loci involved in bacterial virulence [J].
Odenbreit, S ;
Till, M ;
Haas, R .
MOLECULAR MICROBIOLOGY, 1996, 20 (02) :361-373