FIELD: biotechnology.
SUBSTANCE: expression plasmid pFIG-hTNFr vector is constructed, with the physical map shown in Fig.5, size of 6674 bp, containing: a) a sequence encoding a recombinant TNFR1-Fc protein with amino acid sequence shown in Fig. 6b; b) E/P sequence of human T-cell leukemia virus enhancer and elongation factor 1 promoter; c) SP sequence encoding CD33 signal peptide; e) IRES sequence encoding a portion of internal ribosomal landing for encephalomyocarditis virus, and g) GS sequence encoding glutamine synthetase. A method for TNFR1-Fc fusion protein production comprises HEK293 cells culturing, sequentially transformed by introduction of the said pFIG-hTNFr vector and psiRNA-h7SK-miR7 vector with the physical map shown in Fig. 7, in a liquid nutrient medium followed by isolation and purification of TNFR1-Fc protein.
EFFECT: invention provides a recombinant TNFR1-Fc protein with a high activity of tumor necrosis factor inhibition with a high yield.
4 cl, 8 dwg, 7 ex
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