The recombinant plasmids pD1spGBD, STRAIN Escherichia coli-PRODUCER recombinant proteins D1-GBD, recombinant protein D1-GBD AND METHOD FOR PRODUCTION THEREOF, METHOD FOR RESEARCH binding of proteins D1-GBD with antibodies SERA OF PATIENTS, METHOD FOR PRODUCING specific antibodies to proteins D1-GBD

摘要

1. A recombinant plasmid pD1spGBD 4247 bp in size, providing a bifunctional recombinant protein expression D1-GBD, a domain consisting of protein 1 LigA L. interrogans, a spacer of glycine and serine residues and 1,3 β-glyukansvyazyvayuschego Cl domain. thermocellum (GBD), containing! artificial bacterial operon chimeric protein comprising! promoter region of the early promoter of bacteriophage T5 (7-87 bp)! recombinant gene (bp 115-970), encoding a chimeric protein D1-GBD,! untranslated region of a bacterial operon transcription terminator (993-1089 bp); ! bacterial operon bla (4042-3182 bp complementary strand) encoding the beta-lactamase, which is a selectable marker for selection of clones of E. coli transformants by the method of counter-selection; ! bacterial replication initiation region type ColE 1 (2419 bp). ! 2. The strain Escherichia coli M15 [pREP4, pD1spGBD], obtained by transformation of strain Escherichia coli M15 / pREP4 plasmid pD1spGBD, deposited in the Russian National Collection of Industrial Microorganisms FSUE GosNIIgenetika under number B-9899 - D1-GBD producing a recombinant protein consisting of domain 1 LigA protein L. Interrogans, spacer of glycine and serine residues and 1,3 β-glyukansvyazyvayuschego Cl domain. thermocellum (GBD). ! 3. A process for preparing D1-GBD recombinant protein comprising growing cells of the strain Escherichia coli M15 [pREP4, pDlspGBD], obtaining a supernatant containing protein, immobilizing the protein by adding to the supernatant suspension 1,3-β-glyukansoderzhaschego sorbent - zymosan A, incubating, laundering sorbent contacted with a protein immobilized on zymosan concentrated protein A column; removal of the protein from the sorbent, conducting purification to obtain the target protein. ! 4. The recombinant