RECOMBINANT PLASMID DNA pET40CmAP/CGL ENCODING HYBRID BIFUNCTIONAL POLYPEPTIDE CmAP/CGL WITH PROPERTIES OF HIGHLY ACTIVE ALKALINE PHOSPHATASE CmAP AND GALACTOSE SPECIFIC LECTIN CGL, RECOMBINANT STRAIN OF E coli Rosetta (DE3)/pET40CmAP/CGL - PRODUCER OF HYBRID BIFUNCTIONAL POLYPEPTIDE CmAP/CGL AND METHOD OF ITS PREPARATION

摘要

FIELD: biotechnology.SUBSTANCE: invention is a recombinant plasmid pET40CmAP/CGL determining the synthesis of hybrid bifunctional polypeptide CmAP/CGL with the properties of highly active alkaline phosphatase of marine bacterium Cobetia marina (CmAP) and galactose specific lectin of mussels Crenomytilus grayanus (CGL). The plasmid comprises NcoI/SalI - fragment of plasmid pET-40b(+) (Novagen) and the DNA fragment of 2028 base pairs, which is a chimeric gene encoding structural genes of the alkaline phosphatase CmAP and galactose specific lectin CGL, interconnected by polynucleotide encoding the amino acid sequence of the linker (G4S)3EL and enterokinase site D4K. Also a recombinant strain of E. coli Rosetta(DE3)/pET40CmAP/CGL is provided, transformed by the indicated plasmid, - producer of the hybrid bifunctional protein CmAP/CGL, comprising the amino acid sequences of recombinant alkaline phosphatase CmAP and galactose specific lectin CGL. The method of obtaining the hybrid bifunctional protein CmAP/CGL is proposed.EFFECT: invention enables to obtain highly purified preparation of the hybrid protein with the properties of recombinant highly active alkaline phosphatase CmAP and galactose specific lectin CGL, and can be used to detect differences in glycosylation of oncofetal antigens.3 cl, 2 dwg, 3 ex