Identification of Salmonella typhi genes expressed within macrophages by selective capture of transcribed sequences (SCOTS)

被引:83
作者
Daigle, F [1 ]
Graham, JE [1 ]
Curtiss, R [1 ]
机构
[1] Washington Univ, Dept Biol, St Louis, MO 63130 USA
关键词
D O I
10.1046/j.1365-2958.2001.02593.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Salmonella enterica serovar Typhi (S. typhi) is a human-restricted pathogen which causes typhoid fever. Relatively little is known about S. typhi host interaction as animal models of this disease are severely limited by the lack of virulence of S. typhi in other hosts. The virulence of other Salmonella serovars in animal models is dependent on the abilities of these bacteria to survive within host macrophages. We have used selective capture of transcribed sequences (SCOTS) to identify S. typhi genes expressed during growth in human macrophages. This positive cDNA selection technique identified 28 distinct clones representing previously identified as well as novel, uncharacterized and hypothetical gene sequences that are expressed intracellularly. Transcripts for the Vi capsular antigen and genes whose products are involved in stress responses and nutrient acquisition were obtained from intracellular bacteria using SCOTS. Most of these clones are present in the S. typhimurium genome and are also expressed in murine macrophages. Nineteen of these gene sequences were disrupted insertionally in S. typhi, and most of the resulting mutants exhibited a lower level of survival within macrophages compared with the wild-type parent strain. Mutant strains, transformed with a copy of a wild-type gene, exhibited a macrophage survival level similar to that of the parental strain.
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收藏
页码:1211 / 1222
页数:12
相关论文
共 44 条
[1]   Virulence of Salmonella enterica serotype Enteritidis aflagellate and afimbriate mutants in a day-old chick model [J].
Allen-Vercoe, E ;
Sayers, AR ;
Woodward, MJ .
EPIDEMIOLOGY AND INFECTION, 1999, 122 (03) :395-402
[2]   SPACIOUS PHAGOSOME FORMATION WITHIN MOUSE MACROPHAGES CORRELATES WITH SALMONELLA SEROTYPE PATHOGENICITY AND HOST SUSCEPTIBILITY [J].
ALPUCHEARANDA, CM ;
BERTHIAUME, EP ;
MOCK, B ;
SWANSON, JA ;
MILLER, SI .
INFECTION AND IMMUNITY, 1995, 63 (11) :4456-4462
[3]  
ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999
[4]   THE DPS PROMOTER IS ACTIVATED BY OXYR DURING GROWTH AND BY IHF AND A SIGMA(S) IN STATIONARY-PHASE [J].
ALTUVIA, S ;
ALMIRON, M ;
HUISMAN, G ;
KOLTER, R ;
STORZ, G .
MOLECULAR MICROBIOLOGY, 1994, 13 (02) :265-272
[5]   Identification of a new iron regulated locus of Salmonella typhi [J].
Baumler, AJ ;
Tsolis, RM ;
vanderVelden, AWM ;
Stojiljkovic, I ;
Anic, S ;
Heffron, F .
GENE, 1996, 183 (1-2) :207-213
[6]  
BAUMLER AJ, 1994, INFECT IMMUN, V62, P1623
[7]   Analysis of the SOS response in Salmonella enterica serovar Typhimurium using RNA fingerprinting by arbitrarily primed PCR [J].
Benson, NR ;
Wong, RMY ;
McClelland, M .
JOURNAL OF BACTERIOLOGY, 2000, 182 (12) :3490-3497
[8]   A COMPLEMENTATION ANALYSIS OF RESTRICTION AND MODIFICATION OF DNA IN ESCHERICHIA COLI [J].
BOYER, HW ;
ROULLAND.D .
JOURNAL OF MOLECULAR BIOLOGY, 1969, 41 (03) :459-&
[9]   SlyA, a transcriptional regulator of Salmonella typhimurium, is required for resistance to oxidative stress and is expressed in the intracellular environment of macrophages [J].
Buchmeier, N ;
Bossie, S ;
Chen, CY ;
Fang, FC ;
Guiney, DG ;
Libby, SJ .
INFECTION AND IMMUNITY, 1997, 65 (09) :3725-3730
[10]   INDUCTION OF SALMONELLA STRESS PROTEINS UPON INFECTION OF MACROPHAGES [J].
BUCHMEIER, NA ;
HEFFRON, F .
SCIENCE, 1990, 248 (4956) :730-732