RECOMBINANT PLASMID DNA PES3-7 ENCODING PEPTIDE WITH SEQUENCE OF HUMAN GRANULOCYTE COLONY-STIMULATING FACTOR (G-CSF) AND STRAIN ESCHERICHIA COLI BL21(DE3)/PES3-7 AS PRODUCER OF HUMAN RECOMBINANT G-CSF

摘要

FIELD: genetic engineering, in particular production of human granulocyte colony-stimulating factor.;SUBSTANCE: Recombinant plasmid DNA pES3-7 with molecular weight of 3.63 MDa (5907 b.p.) is constructed. Said DNA consists DNA Ndel/Notl-fragment containing sequence of recombinant G-CSF artificial gene, β-lactamase gene; and plasmid pET22b(+) DNA Ndel/Notl-fragment containing promoter and terminator of T-RNA-polymerase transcription, amplifier of 17 phage 10 gene translation. Plasmid pES3-7 contains as genetic marker β-lactamase gene which determines resistance of E.coli cells transformed with plasmid pES3-7 to ampicillin, and unique restriction endonuclease recognition sites existing on the next distance to the right from Ndel-site: Xbal - 38 b.p.; Hpal - 1332 b.p.; Pstl - 4065 b.p.; Pvul - 4190 b.p.; Xhol - 5363 b.p. Obtained plasmid is used in transformation of Escherichia coli cells to produce strain E.coli BL21(DE3)/pES3-7 as subproducer of recombinant G-CSF. Method of present invention makes in possible to produce recombinant G-CSF with high yield (20-30 % based on total cell protein content).;EFFECT: simplified method for production of recombinant G-CSF with high yield.;2 cl, 2 dwg, 2 ex