RECOMBINANT PLASMID DNA ENCODING SYNTHESIS, METHOD FOR PREPARING AND PREPARATION OF HUMAN RECOMBINANT GAMMA-INTERFERON

摘要

FIELD: biotechnology, molecular biology, genetic engineering, pharmacy. SUBSTANCE: invention relates to method for preparing and preparation of human recombinant gamma-interferon. Constructed recombinant plasmid pGIF 315 comprises promoters providing high constitutive expression of gene in cells E. coli encoding gamma-interferon with size 144 pairs of nitrogen bases with molecular mass 16.9 kDa. Method for preparing interferon involves culturing strain E. coli transformed with plasmid pGIF 315 in nutrient medium without deficiency of any components or addition of any inducing agents for synthesis of gamma-interferon. After lysis of cells precipitate is washed out with 1-3 M urea solutions and dissolved in 6-8 M urea solution. Prepared solution is applied on cation-exchange sorbent and elution with urea-free buffer is carried out that provides renaturation of gamma-interferon. Preparation of human recombinant gamma-interferon comprises additionally biologically inert low-molecular filling agent and salt. Invention provides creation of plasmid resulting to high yield of gamma-interferon. Also, invention relates to creation of medicinal preparations of human gamma-interferon and recombinant plasmid DNA encoding its synthesis. EFFECT: improved preparing method. 6 cl, 1 tbl