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ISSN 1995-5537

Cover 5 2010

"Biotechnology" journal Vol. 3, No. 5, 2010
Р. 50-56, Bibliography 17, Ukrainian.
Universal Decimal classification: 616–002.5+577.27


T. A. Redchuk, N. V. Korotkevich, A. A. Kaberniuk, O. S. Oliinyk, A. J. Labyntsev, S. I. Romaniuk, D. V. Kolibo, S. V. Komisarenko

Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, Kyiv

Proteins of Mycobacterium bovis MPB63 and MPB83 are perspective candidates for development of serological test-systems to detect tuberculosis. We have obtained expression vector carrying sequences of genes mpb63 and mpb83 in single open reading frame and the corresponding chimeric protein MPB63–MPB83, that can be purified by metal-affinity chromatography.

Experiments using sera from rabbits immunized with antigens MPB63 and MPB83 showed not only match the antigenic structure of the fused protein and its constituent parts, but also the opportunity to achieve a higher level of signal in ELISA using chimeric construction, probably due to the better adsorption properties.

Also, we have determined the level of antibodies against the fusion proteins MPB63–MPB83 in sera of healthy, immunized with BCG, experimentally infected with different types of mycobacteria (M. bovis, M. intracellulare, M. scrofulaceum, M. fortuitum) and leukaemic cattle. The results of these experiments testified that only the animals infected with M. bovis showed high levels of antibodies against the fusion protein MPB63–MPB83. Thereby, we have obtained genetically fused protein MPB63-MPB83 which could be used to develop ELISA test systems for diagnosis of bovine tuberculosis.

Key words: recombinant fusion protein, MPB83, MPB63, Mycobacterium bovis, antigenic properties, immunodiagnostics, tuberculosis, cattle.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008