RECOMBINANT PLASMID DNA pMALTEV-legumain, ENCODING POLYPEPTIDE HAVING ANTIGENIC PROPERTIES OF PROTEIN LEGUMAIN Opisthorchis felineus, AND STRAIN E.coli BL 21(DE3)pLysS-pMALTEV-legumain - PRODUCENT OF RECOMBINANT POLYPEPTIDE HAVING ANTIGENIC PROPERTIES OF PROTEIN LEGUMAIN Opisthorchis felineus Russian patent published in 2013 - IPC C12N15/00 C12N1/21 

FIELD: biotechnologies.

SUBSTANCE: invention is recombinant plasmid DNA pMALTEV-legumain, with molecular weight of 4.78 megadalton and size of 7839 bp, coding a polypeptide having antigenic properties of protein legumain Opisthorchis felineus, and containing a fragment of a vector plasmid pMALTEV, including a Ptac promotor; a lac-operator sequence; a sequence of a gene malE, coding a maltose-binding protein; a combination of terminators of transcription rrnB T1T2 E.coli; a gene of b-lactamase and a section of ori (pBR322) replication initiation; complementary DNA of gene legumain O.felineus without a signal peptide flanked with sites of restriction BamHI and Hindlll; a hybrid promotor Ptac; a lac-operator sequence for amplification of lac-repressor binding; terminators of transcription of gene rrnB E.coli (t1 and t2); as a genetic marker - a gene of B-lactamase (amp^R), which determines resistance of E.coli cells transformed with plasmid pMALTEV-legumain to ampicillin antibiotics; a nucleotide sequence, which codes MBP (maltose-binding protein), being a part of a hybrid sequence of a recombinant protein MBP-legumain and making it possible to separate a recombinant polypeptide by methods of affine chromatography; unique sites of endonucleases of restriction, having the following coordinates: BamHI (2668), EcoRI (2675), StuI (2685), Sail (2691), Spel (2704), NotI (2711), Xbal (2725), PstI (2737), Xhol (2740), SphI (2750), Kpnl (2756), Hindlll (2758). The invention also relates to a strain E.coli BL21(DE3)pLysS-pMALTEV-legumain, produced with the help of the recombinant plasmid DNA pMALTEV-legumain, deposited in the collection of microorganism cultures in the Federal Budget Institution of Science State Science Centre of Virusology and Biotechnology "Vector" of Federal Service for Oversight of Consumer Protection and Welfare under the number B-1253. The invention makes it possible to produce a recombinant hybrid polypeptide with antigenic properties of protein legumain O.felineus, providing for high specificity to antibodies of a parasitic infection O.felineus.

EFFECT: high-sensitive test-systems with application of the specified polypeptide.

2 cl, 6 dwg, 5 ex