Identification of genes encoding exported Mycobacterium tuberculosis proteins using a Tn552'phoA in vitro transposition system

Miriam Braunstein, Thomas J. Griffin IV, Jordan I. Kriakov, Sarah T. Friedman, Nigel D.F. Grindley, William R. Jacobs

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Secreted and cell envelope-associated proteins are important to both Mycobacterium tuberculosis pathogenesis and the generation of protective immunity to M. tuberculosis. We used an in vitro Tn552'phoA transposition system to identify exported proteins of M. tuberculosis. The system is simple and efficient, and the transposon inserts randomly into target DNA. M. tuberculosis genomic libraries were targeted with Tn552'phoA transposons, and these libraries were screened in M. smegmatis for active PhoA translational fusions. Thirty-two different M. tuberculosis open reading frames were identified; eight contain standard signal peptides, six contain lipoprotein signal peptides, and seventeen contain one or more transmembrane domains. Four of these proteins had not yet been assigned as exported proteins in the M. tuberculosis databases. This collection of exported proteins includes factors that are known to participate in the immune response of M. tuberculosis and proteins with homologies, suggesting a role in pathogenesis. Nine of the proteins appear to be unique to mycobacteria and represent promising candidates for factors that participate in protective immunity and virulence. This technology of creating comprehensive fusion libraries should be applicable to other organisms.

Original languageEnglish (US)
Pages (from-to)2732-2740
Number of pages9
JournalJournal of Bacteriology
Volume182
Issue number10
DOIs
StatePublished - May 1 2000

Fingerprint

Mycobacterium tuberculosis
Genes
Proteins
Protein Sorting Signals
Libraries
Immunity
Smegma
Genomic Library
Mycobacterium
In Vitro Techniques
Open Reading Frames
Lipoproteins
Virulence
Databases
Technology
DNA

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Identification of genes encoding exported Mycobacterium tuberculosis proteins using a Tn552'phoA in vitro transposition system. / Braunstein, Miriam; Griffin IV, Thomas J.; Kriakov, Jordan I.; Friedman, Sarah T.; Grindley, Nigel D.F.; Jacobs, William R.

In: Journal of Bacteriology, Vol. 182, No. 10, 01.05.2000, p. 2732-2740.

Research output: Contribution to journalArticle

Braunstein, Miriam ; Griffin IV, Thomas J. ; Kriakov, Jordan I. ; Friedman, Sarah T. ; Grindley, Nigel D.F. ; Jacobs, William R. / Identification of genes encoding exported Mycobacterium tuberculosis proteins using a Tn552'phoA in vitro transposition system. In: Journal of Bacteriology. 2000 ; Vol. 182, No. 10. pp. 2732-2740.
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