| 摘要 |
FIELD: organic chemistry, genetics. ^ SUBSTANCE: invention refers to genetic engineering, specifically to production of proinsulin Glargin and can be applied for production of medicines of new generation used for insulin-dependent pancreatic diabetes. Designed recombinant plasmid DNA pGG-1 codes hybrid protein with amino acid sequence of proinsulin Glargin, where sequence of domain B of protein A Staphylococcus aureus is connected through peptide linker His6GlySerArg with amino acid sequence of proinsulin Glargin having molecular weight 3,3 MDa (5051 "."), containing HindIII/BamHI-fragment of plasmide pPINS07, including hybride tac-promoter, gene of beta-lactamase ("bla"), replication origin area (ori), terminator of ribosomal operon E. coli transcription, nucleotide sequence coding amino acid sequence of domain B of protein A Staphylococcus aureus, connected to nucleotide sequence coding peptide His6GlySerArg, HindIII/BamHI-fragment (271 "."), coding proinsulin Glargin; unique restriction sites with following coordinates: EcoRI - 1, NcoI - 217, BamHI - 221, PstI - 342 and 417, HindIII - 492, SalGI - 4513, ClaI - 4792. Bacteria Escherichia coli BL21 are transformed with plasmide pGG-1 that resulted in strains Escherichia coli BGG18, producer of hybride polypeptide containing amino acid sequence of proinsulin Glargin. Invention enables to produce insulin Glargin with high yield and applying simplified technology. ^ EFFECT: production of insulin Glargin with high yield and applying simplified technology. ^ 2 cl, 4 dwg, 4 ex |
