Expression of the Mycobacterium tuberculosis 19-kilodalton antigen in Mycobacterium smegmatis: Immunological analysis and evidence of glycosylation

T. Garbe*, D. Harris, M. Vordermeier, R. Lathigra, J. Ivanyi, D. Young

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

The gene encoding a 19-kDa antigen from Mycobacterium tuberculosis was expressed as a recombinant protein in the rapid-growing species Mycobacterium smegmatis. The recombinant antigen was expressed at a level approximately ninefold higher than in M. tuberculosis and, like the native antigen, was found in the pellet fraction after high-speed centrifugation of bacterial extracts. The 19-kDa antigen in crude bacterial extracts, and the purified recombinant antigen, bound strongly to concanavalin A, indicating the possibility of posttranslational glycosylation. The recombinant antigen stimulated T-cell proliferation in vitro when added to assays either in the form of whole recombinant bacteria or as a purified protein. Homologous expression of mycobacterial antigens in a rapid-growing mycobacterial host may be particularly useful for the immunological characterization of proteins which are subject to posttranslational modification.

Original languageEnglish
Pages (from-to)260-267
Number of pages8
JournalInfection and Immunity
Volume61
Issue number1
DOIs
Publication statusPublished - 01 Jan 1993

Keywords

  • Amino Acid Sequence
  • Animals
  • Antigens, Bacterial/biosynthesis
  • Blotting, Western
  • Concanavalin A/metabolism
  • Dose-Response Relationship, Immunologic
  • Electrophoresis, Polyacrylamide Gel
  • Glycosylation
  • Lymphocyte Activation/immunology
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Mycobacterium/metabolism
  • Mycobacterium tuberculosis/immunology
  • Protein Processing, Post-Translational
  • Recombinant Proteins/biosynthesis
  • T-Lymphocytes/cytology

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