RECOMBINANT PLASMID DNA PUABC 22 ENCODING MODIFIED ACTIVATOR OF URICASE TYPE PLASMINOGEN, NONTRANSLATED DNA-ELEMENT - ARTIFICIAL INTERGENE SEQUENCE MGP-14 AND STRAIN OF BACTERIUM ESCHERICHIA COLI - PRODUCER OF MODIFIED ACTIVATOR OF URICASE TYPE PLASMINOGEN

摘要

FIELD: genetic engineering, biotechnology. SUBSTANCE: invention relates to biotechnology of preparing highly productive strains of Escherichia coli - producers of human recombinant proteins used in the modern medicine as thrombolytic agents. New recombinant plasmid DNA pUABC 22 encoding the modified activator of plasminogen of uricase type (mAPUT) has a size 4.5 t. n. p. and has the following components: XmnI-Bam HI-fragment of DNA of plasmid pUABC 22 (size is 2.99 t. n. p.); Bam HI-KpnI-fragment of DNA (size is 0.177 t. n. p.) containing the site of encoding region of gene mAPUT and corresponding to 12 C-terminal amino acids of mAPUT and nontranslated element - an artificial intergene sequence MGP-14; KpnI-SacI-fragment of DNA (size is 0.57 t. n. p.) containing gene tRNA Arg; SacI-XnmI-fragment of DNA of vector pUC 19 (size is 0.79 t. n. p.); site of replication initiation and promoter-operator region of lac-operon; genetic markers; gene determining the resistance to ampicillin. Nontranslated DNA-elements - an artificial intergene sequence MGP-14 and gene tRNA Arg enhance the total stability of the expression system of mAPUT in Escherichia coli cells and enhance its translating activity with respect to eucaryotic genes. The new strain VKPM B-7666 produces 3.0 g cellular biomass/1 l of cultural fluid and produces mAPUT in an proenzyme form mainly (molecular mass is 43 kDa) at amount 200 000-250 000 U/1 g of biomass. EFFECT: new strain indicated above, increased yield of the plasminogen activator. 4 cl, 4 dwg, 4 ex