FIELD: biotechnology.
SUBSTANCE: invention relates to microbiological industry, biotechnology, genetic and protein engineering and concerns recombinant plasmid DNA pET19b-SAV, which provides synthesis of full-size protein streptavidin, having molecular weight 4 MDa and size 6126 base pairs and containing, in accordance with physical and genetic map of plasmid, restriction sites of NcoI, XhoI, NcoI-XhoI DNA fragment of plasmid pET19b with length of 5643 base pairs, including T7lac promoter, lacI protein of lac protein repressor protein, gene β-lactamase, determining ampicillin resistance during E. coli transformation, a replication initiation ori; DNA fragment of NcoI-XhoI with size of 483 base pairs, which is a sequence coding full-length streptavidin without leader peptide SEQ ID No. 1; as well as Escherichia coli bacterium strain - producer of full-size streptavidin produced by transformation of cell culture Escherichia coli BL21-Codon Plus (DE3) plasmid DNA pET19b-SAV, having nucleotide sequence SEQ ID No. 1 protein of full-length streptavidin.
EFFECT: invention enables to obtain a recombinant Escherichia coli strain capable of synthesising streptavidin with complete biological activity into a cytoplasmic solution, which simplifies the procedure for obtaining highly purified protein (single-step affinity chromatography) compared to production of streptavidin from periplasmic, as well as from insoluble fractions, requiring several stages of purification.
2 cl, 4 dwg, 3 ex
