Expression, immunochemical characterization and localization of the Mycobacterium tuberculosis protein p27.
Rv2108 is a gene of the PPE family of Mycobacterium tuberculosis specific for this bacterial complex and that may encode a putative protein p27. This gene was amplified, inserted into bacterial vectors, sequenced, and expressed as a recombinant protein. Specific antibodies to this protein were generated and used for immunochemical characterization and cellular localization. Mass spectrometric analysis of the expressed protein revealed a molecule that corresponded to the p27 putative protein. The expressed protein was immunologically active, and reacted with antibodies from tuberculosis patient sera. Specific immunoblot analysis confirmed the presence of the p27 antigen in Mycobacterium bovis BCG strain and in human clinical isolates of M. tuberculosis, but not in other mycobacteria tested. Western blot and immunoelectron microscopic analysis of BCG strain indicated that the p27 protein is localized in the membrane of the cell. The specific expression of the p27 protein in the M. tuberculosis complex could provide a novel specific complimentary diagnostic test for the presence of and infection with M. tuberculosis.
Le Moigne
V
V
Laboratoire Universitaire de Microbiologie Appliquée de Quimper (LUMAQ), UBO, 2 rue de l'Université, Quimper 29334 cedex, France.
Robreau
G
G
Borot
C
C
Guesdon
J-L
JL
Mahana
W
W
D000907:Antibodies, Bacterial / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial / Q000276:immunology*; D006801:Humans; D007158:Immunologic Techniques; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D011994:Recombinant Proteins / Q000032:analysis; D014376:Tuberculosis / Q000276:immunology*
D000907:Antibodies, Bacterial; D008074:Lipoproteins; C110121:P27 protein, Mycobacterium; D011994:Recombinant Proteins