The recombinant plasmids pD4spGBD, STRAIN Escherichia coli - PRODUCER recombinant protein D4-GBD, recombinant protein D4-GBD AND METHOD FOR PRODUCTION THEREOF, METHOD FOR RESEARCH protein binding D4-GBD with antibodies SERA OF PATIENTS, METHOD FOR PRODUCING specific antibodies to the protein D4-GBD

摘要

1. A recombinant plasmid pD4spGBD size 4271 bp, providing expression of the recombinant bifunctional protein D4-GBD, consisting of 4 protein domain 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 (115-994 bp) encoding a chimeric protein D4-GBD,! untranslated region of a bacterial operon transcription terminator (1017-1113 bp); ! bacterial operon bla (4066-3206 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 ColE1 (2443 bp). ! 2. The strain Escherichia coli M15 [pREP4, pD4spGBD], obtained by transformation of strain Escherichia coli M15 / pREP4 plasmid pD4spGBD, deposited in the Russian National Collection of Industrial Microorganisms FSUE GosNIIgenetika under number B-9901 - D4-GBD producing a recombinant protein consisting of domain 4 LigA protein L. Interrogans, spacer of glycine and serine residues and 1,3 (3-Cl glyukansvyazyvayuschego domain. thermocellum (GBD).! 3. A method of producing recombinant protein D4-GBD, comprising growing cells of the strain Escherichia coli M15 [pREP4, pD4spGBD], obtaining the supernatant containing the protein, protein immobilization way adding to the supernatant suspension 1,3-β-glyukansoderzhaschego sorbent - zymosan A, incubation, laundering sorbent contacted with a protein concentration of zymosan immobilized on protein A column, protein removal from the sorbent, conducting purification to obtain the target recombinant protein 4.!