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Preparation and characterization of recombinant protein containing immunomimetic peptide of benzo(a)pyrene

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Abstract

Two recombinant plasmids were constructed. The first plasmid contained a hybrid gene composed of the sequences that encode immunomimetic peptide of benzo(a)pyrene, protein pIII of bacteriophage M13, and cellulose binding domain. The second plasmid contained a hybrid gene composed of the sequences of signal peptide of protein pIII of bacteriophage M13, immunomimetic peptide of benzo(a)pyrene, protein pIII of bacteriophage M13, and cellulose binding domain. The obtained recombinant plasmids were used to create producer strains of a chimeric protein containing immunomimetic peptide of benzo(a)pyrene based on the strain E. coli M15. The recombinant protein was only shown to accumulate in producer cells under the induction of the expression of the hybrid gene with the signal peptide of protein pIII of bacteriophage. This chimeric protein was expressed in mature (without the signal peptide) and unprocessed (with the signal peptide) forms. Western-blot analysis showed that only the mature form specifically reacted with the B2 monoclonal antibodies against benzo(a)pyrene. Thus, recombinant protein containing immunomimetic peptide of benzo(a)pyrene has been expressed, purified, and characterized.

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Correspondence to S. V. Apalko.

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Original Russian Text © S.V. Apalko, V.G. Lunin, M.L. Filipenko, V.A. Matveeva, A.M. Lyashchuk, N. V. Lavrova, E.A. Sherina, A.V. Averjanov, M.V. Kostyanko, A.N. Glushkov, 2011, published in Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 2011, No. 2, pp. 24–29.

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Apalko, S.V., Lunin, V.G., Filipenko, M.L. et al. Preparation and characterization of recombinant protein containing immunomimetic peptide of benzo(a)pyrene. Mol. Genet. Microbiol. Virol. 26, 74–81 (2011). https://doi.org/10.3103/S0891416811020029

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  • DOI: https://doi.org/10.3103/S0891416811020029

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