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PMID:10781540

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Citation

Braunstein, M, Griffin TJ, IV, Kriakov, JI, Friedman, ST, Grindley, ND and Jacobs, WR Jr (2000) Identification of genes encoding exported Mycobacterium tuberculosis proteins using a Tn552'phoA in vitro transposition system. J. Bacteriol. 182:2732-40

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.

Links

PubMed PMC101980

Keywords

Alkaline Phosphatase/metabolism; Amino Acid Sequence; Artificial Gene Fusion; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Biological Transport; Cosmids; Cyclin-Dependent Kinases/genetics; DNA Transposable Elements; DNA, Bacterial; Genes, Bacterial; Genomic Library; Molecular Sequence Data; Mutagenesis, Insertional; Mycobacterium tuberculosis/genetics; Mycobacterium tuberculosis/metabolism; Plasmids; Replication Origin

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MYCTU:A85A

part_of

GO:0005576: extracellular region

ECO:0000316: genetic interaction evidence used in manual assertion

C

Seeded From UniProt

Missing: with/from

MYCTU:PSTS1

part_of

GO:0005576: extracellular region

ECO:0000316: genetic interaction evidence used in manual assertion

C

Seeded From UniProt

Missing: with/from

See also

References

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