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

France Daigle, James E. Graham, Roy Curtiss

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1211-1222
Number of pages12
JournalMolecular Microbiology
Volume41
Issue number5
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Salmonella typhi
Macrophages
Genes
Virulence
Clone Cells
Bacteria
Animal Disease Models
Typhoid Fever
Salmonella
Animal Models
Complementary DNA
Serogroup
Genome
Antigens
Survival
Growth

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Identification of Salmonella typhi genes expressed within macrophages by selective capture of transcribed sequences (SCOTS). / Daigle, France; Graham, James E.; Curtiss, Roy.

In: Molecular Microbiology, Vol. 41, No. 5, 2001, p. 1211-1222.

Research output: Contribution to journalArticle

Daigle, France ; Graham, James E. ; Curtiss, Roy. / Identification of Salmonella typhi genes expressed within macrophages by selective capture of transcribed sequences (SCOTS). In: Molecular Microbiology. 2001 ; Vol. 41, No. 5. pp. 1211-1222.
@article{64e7485856d14b23997efe5fd41de014,
title = "Identification of Salmonella typhi genes expressed within macrophages by selective capture of transcribed sequences (SCOTS)",
abstract = "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.",
author = "France Daigle and Graham, {James E.} and Roy Curtiss",
year = "2001",
doi = "10.1046/j.1365-2958.2001.02593.x",
language = "English (US)",
volume = "41",
pages = "1211--1222",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "5",

}

TY - JOUR

T1 - Identification of Salmonella typhi genes expressed within macrophages by selective capture of transcribed sequences (SCOTS)

AU - Daigle, France

AU - Graham, James E.

AU - Curtiss, Roy

PY - 2001

Y1 - 2001

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0035788804&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035788804&partnerID=8YFLogxK

U2 - 10.1046/j.1365-2958.2001.02593.x

DO - 10.1046/j.1365-2958.2001.02593.x

M3 - Article

C2 - 11555299

AN - SCOPUS:0035788804

VL - 41

SP - 1211

EP - 1222

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 5

ER -