FIELD: chemistry.
SUBSTANCE: inventions relate to vector construction, Escherichia coli strain, including such vector construction, and method for obtaining recombinant analgesic peptide PT1. Vector construction is obtained on the basis of vector pTWIN1 and contains gene of chimeric protein DnaB-PT1, which consists of chitin-binding domain from Bacillus circulans, intein DnaB from Synechocystis sp. and analgesic peptide PT1. Characterised method for obtaining includes carrying out controlled expression of gene of chimeric protein DnaB-PT1 in composition of characterised vector construction in E. coli strain-producent ER2566 with application of isopropyl-β-D-1-thiogalactopyranoside as inductor. After that lysis of cells, separation of soluble components, purification of chimeric protein DnaB-PT1 by means of affinity chromatography on chitin sorbent, autocatalytic cleavage of chimeric protein DnaB-PT1 and purification of peptide PT1 by means of reverse phase chromatography are performed.
EFFECT: claimed group of inventions makes it possible to obtain analgesic peptide with high output for application for medical purposes.
3 cl, 8 dwg, 6 ex
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Number |
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