Dissection of Mycobacterium tuberculosis Antigens Using Recombinant DNA

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 82; no. 9; pp. 2583 - 2587
• Main Authors: Young, Richard A., Bloom, Barry R., Grosskinsky, Clemens M., Ivanyi, Juraj, Thomas, Darcy, Davis, Ronald W.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.05.1985; National Acad Sciences
• Subjects: Antibodies; Antibodies, Monoclonal - immunology; Antigens; Antigens, Bacterial - genetics; Bacteriology; Bacteriophages; Biological and medical sciences; Biotechnology; Cloning, Molecular; Coliphages; DNA; DNA, Bacterial - genetics; DNA, Recombinant; Epitopes - genetics; Fundamental and applied biological sciences. Psychology; Genomes; Health. Pharmaceutical industry; Industrial applications and implications. Economical aspects; Libraries; Microbiology; Monoclonal antibodies; Mycobacterium tuberculosis; Mycobacterium tuberculosis - genetics; Mycobacterium tuberculosis - immunology; Pathogens; Production of active biomolecules; Recombinant DNA; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies; Vaccins; Diagnostic; Vaccine; Mycobacterial infection; Monoclonal antibody; Epidemiology; Immunological method; Infection; Antigen; Proteins; Tuberculosis; Gene; Mycobacterium tuberculosis; Mycobacteriales; Test; Serological method; Preparation; Bacteriosis; Mycobacteriaceae; Bacteria; Isolation; Actinomycetes; Molecular cloning
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.82.9.2583

A recombinant DNA strategy has been used systematically to survey the Mycobacterium tuberculosis genome for sequences that encode specific antigens detected by monoclonal antibodies. M. tuberculosis genomic DNA fragments with randomly generated endpoints were used to construct a large λ gt11 recombinant DNA expression library. Sufficient numbers of recombinants were produced to contain inserts whose endpoints occur at nearly every base pair in the pathogen genome. Protein antigens specified by linear segments of pathogen DNA and produced by the recombinant phage of Escherichia coli were screened with monoclonal anti-body probes. This approach was coupled with an improved detection method for gene isolation using antibodies to clonally isolate DNA sequences that specify polypeptide components of M. tuberculosis. The methodology described here, which is applicable to other pathogens, offers possibilities for the development of more sensitive and specific immunodiagnostic and sero-epidemiological tests for tuberculosis and, ultimately, for the development of more effective vaccines.