FIELD: biotechnology.
SUBSTANCE: invention refers to the recombinant production of human proteins and can be used in order to prepare human orexin A in the Escherichia coli cells. Production method consists in culturing the cells of the producer strain Escherichia coli BL21(DE3)pET151/D-TOPO, which is obtained by means of transformation of plasmid DNA pET151/D-TOPO, which contains the codon optimized human orexin A gene, of the Escherichia coli cells BL21 (DE3) using the inducer of the target protein expression. Cells are then lysed, the inclusion bodies are washed with 0.2 M sodium deoxycholate, the inclusion bodies are dissolved in the 8 M urea solution, orexin A is refolded in 0.1M Tris pH 8.0 buffer, 0.2 mM EDTA with 0.5 M L-arginine and chromatographic purification of the resulting protein solution on the S-Sepharose column and on the S-100 column is carried out.
EFFECT: invention allows to obtain the highly purified physiologically active recombinant human orexin A.
3 cl, 3 dwg, 2 tbl, 5 ex
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