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
[1]   A genome-scale analysis for identification of genes required for growth or survival of Haemophilus influenzae [J].
Akerley, BJ ;
Rubin, EJ ;
Novick, VL ;
Amaya, K ;
Judson, N ;
Mekalanos, JJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (02) :966-971
[2]   Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori [J].
Alm, RA ;
Ling, LSL ;
Moir, DT ;
King, BL ;
Brown, ED ;
Doig, PC ;
Smith, DR ;
Noonan, B ;
Guild, BC ;
deJonge, BL ;
Carmel, G ;
Tummino, PJ ;
Caruso, A ;
Uria-Nickelsen, M ;
Mills, DM ;
Ives, C ;
Gibson, R ;
Merberg, D ;
Mills, SD ;
Jiang, Q ;
Taylor, DE ;
Vovis, GF ;
Trost, TJ .
NATURE, 1999, 397 (6715) :176-180
[3]   Comparative genomics of Helicobacter pylori:: Analysis of the outer membrane protein families [J].
Alm, RA ;
Bina, J ;
Andrews, BM ;
Doig, P ;
Hancock, REW ;
Trust, TJ .
INFECTION AND IMMUNITY, 2000, 68 (07) :4155-4168
[4]  
AUSEBEL F, 1997, SHORT PROTOCOLS MOL
[5]  
Ausubel FM, 1995, CURRENT PROTOCOLS MO
[6]   Molecular characterization of two-component systems of Helicobactcr pylori [J].
Beier, D ;
Frank, R .
JOURNAL OF BACTERIOLOGY, 2000, 182 (08) :2068-2076
[7]   A simple in vitro Tn7-based transposition system with low target site selectivity for genome and gene analysis [J].
Biery, MC ;
Stewart, FJ ;
Stellwagen, AE ;
Raleigh, EA ;
Craig, NL .
NUCLEIC ACIDS RESEARCH, 2000, 28 (05) :1067-1077
[8]   Identification of virulence genes of Helicobacter pylori by random insertion mutagenesis [J].
Bijlsma, JJE ;
Vandenbroucke-Grauls, CMJE ;
Phadnis, SH ;
Kusters, JG .
INFECTION AND IMMUNITY, 1999, 67 (05) :2433-2440
[9]   Identification of loci essential for the growth of Helicobacter pylori under acidic conditions [J].
Bijlsma, JJE ;
Lie-A-Ling, M ;
Nootenboom, IC ;
Vandenbroucke-Grauls, CMJE ;
Kusters, JG .
JOURNAL OF INFECTIOUS DISEASES, 2000, 182 (05) :1566-1569
[10]   mraW, an essential gene at the dcw cluster of Escherichia coli codes for a cytoplasmic protein with methyltransferase activity [J].
Carrión, M ;
Gómez, MJ ;
Merchante-Schubert, R ;
Dongarrá, S ;
Ayala, JA .
BIOCHIMIE, 1999, 81 (8-9) :879-888