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Cloning and Expression of Protective Antigens of Mycobacterium tuberculosis Ag85B and ESAT-6 in Francisella tularensis 15/10

T. B. Kravchenko*, M. E. Platonov, G. M. Vahrameeva, V. A. Bannov, T. Ju. Kudryavtseva, A. N. Mokrievich, and V. M. Pavlov

State Research Center of Applied Microbiology and Biotechnology, 142279 Obolensk, Moscow Region, Russia; fax: (4967) 36-0061; E-mail: kravchenko@obolensk.org

* To whom correspondence should be addressed.

Received April 3, 2007
The possibility of expression of genes encoding mycobacterial antigens in Francisella tularensis 15/10 vaccine strain cells has been shown for the first time. To obtain stable and effective expression of mycobacterial antigens in the F. tularensis cells, the plasmid vector pPMC1 and hybrid genes consisting of the leader part FL of the F. tularensis membrane protein FopA and structural moieties of the mature protein Ag85B or the fused protein Ag85B-ESAT-6 were constructed. Recombinant strains F. tularensis RVp17 and RVp18 expressing protective mycobacterial antigens in the fused proteins FL-Ag85B and FL-Ag85B-ESAT-6, respectively, were obtained. Expression of the protective mycobacterial antigens in F. tularensis was analyzed using specific antisera to the recombinant proteins Ag85-(His)6 and ESAT-6-(His)6 isolated from Escherichia coli producer strains created on the basis of the pET23b(+) and pET24b(+) vectors. The expression of heterologous protective antigens in F. tularensis 15/10 is promising for creation of live recombinant anti-tuberculosis vaccines on the basis of the tularemia vaccine strain.
KEY WORDS: expression, Ag85B, ESAT-6, Mycobacterium tuberculosis, Francisella tularensis

DOI: 10.1134/S0006297907070073