RECOMBINANT PLASMID DNA pET40CmAP/MBL-T CODING HYBRID BIFUNCTIONAL POLYPEPTIDE CmAP/MBL-T WITH PROPERTIES OF HIGH-ACTIVITY ALKALINE PHOSPHATASE CmAP AND MANNAN-BINDING LECTIN OF C-TYPE MBL-T, RECOMBINANT STRAIN E coli Rosetta(DE3)/pET40CmAP/MBL-T - PRODUCER OF HYBRID BIFUNCTIONAL POLYPEPTIDE CmAP/MBL-T, AND METHOD FOR ITS OBTAINING

摘要

FIELD: biotechnologies.;SUBSTANCE: invention represents plasmid pET40CmAP/MBL-T determining a synthesis of hybrid bifunctional polypeptide CmAP/MBL-T with properties of high-activity alkaline phosphatase of sea bacterium Cobetia marina (CmAP) and mannan-binding lectin of C-type of far-eastern trepang Apostichopus japonicus (MBL-T). The plasmid includes NcoI/SalI - a fragment of plasmid pET-40b(+) (Novagen) and a DNA fragment with size of 2034 pairs of bases, which is a chimeric gene coding structural genes of alkaline phosphatase CmAP and mannan-binding lectin of C-type MBL-T, which are connected to each other with a polynucleotide coding the amino acid sequence of linker (G)₄S(G)₄S(G)₄SEL. The invention describes recombinant strain E.coli Rosetta(DE3)/pET40CmAP/MBL-T transformed by the above plasmid - a producer of hybrid bifunctional protein CmAP/MBL-T. The invention also proposes a method for obtaining hybrid bifunctional protein CmAP/MBL-T using the proposed strain.;EFFECT: invention allows obtaining the specified highly purified preparation of a hybrid protein with properties of recombinant high-activity alkaline phosphatase CmAP and highly specific mannan-binding lectin of C-type MBL-T with regard to carbohydrate components of glycoproteins of cancer cells of uterine cervix and can be used for a high-sensitivity lectin-immunoferment method of early differential diagnostics of cervical cancer.;3 cl, 2 dwg, 3 ex