Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS)

James E. Graham, Josephine E. Clark-Curtiss

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357 Citations (Scopus)

Abstract

A widely applicable, positive cDNA selection method was developed to identify RNAs synthesized by Mycobacterium tuberculosis in response to phagocytosis by cultured human primary macrophages, cDNAs for sigE and sigH (alternative sigma factors), aceA (isocitrate lyase), ponA (class I penicillin-binding protein), pks2 (polyketide synthase), uvrA (UvrABC endonuclease), and ctpV (putative cation transporter) were obtained from macrophage-grown bacteria, cDNAs for ORFs Rv3070, Rv3483c, Rv0903c (encoding a putative bacterial two-component transcriptional activator), and Rv0170 of the mce1 virulence operon also were obtained from phagocytized bacilli, cDNAs for these genomic regions were not obtained from approximately 1,000-fold more bacteria grown in laboratory broth. Methods described here, which have identified M. tuberculosis genes expressed in response to host interaction, will allow the study of gene expression in a variety of microorganisms, including expression resulting from interaction with human tissues in natural disease states.

Original languageEnglish (US)
Pages (from-to)11554-11559
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number20
DOIs
StatePublished - Sep 28 1999
Externally publishedYes

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Mycobacterium tuberculosis
Phagocytosis
Complementary DNA
Macrophages
RNA
Isocitrate Lyase
Polyketide Synthases
Penicillin-Binding Proteins
Bacteria
Sigma Factor
Endonucleases
Operon
Bacillus
Open Reading Frames
Virulence
Cations
Gene Expression
Genes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

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