RU2004104194A - Recombinant plasmid DNA PHINSO5, encode fusion polypeptides comprising human proinsulin CELL ESCHERICHIA COLI, transformed with recombinant plasmid DNA PHINSO5, STRAIN BACTERIA AND ESCHERICHIA COLI JM109 / PHINSO5-PRODUCER HYBRID polypeptide comprising human proinsulin - Google Patents

Claims (3)

1. Recombinant plasmid DNA pHINS05 encoding a hybrid polypeptide with the amino acid sequence of human proinsulin, in which the N-terminal fragment of human gamma interferon is connected via the peptide linker His ₄ GlySerArg with the amino acid sequence of human proinsulin, having a molecular weight of 2.75 MDa, containing BamHI- EcoRI - fragment of plasmid pKK223-3, including the tac transcription promoter, EcoRI-HindIII - fragment with a gene having the nucleotide sequence shown in Fig. 2, and encoding the N-terminal fragment of gamma inter human feron, peptide linker His ₄ GlySerArg and amino acid sequence of human proinsulin, HindIII-EheI - fragment of plasmid pKK223-3, including the transcription terminator of the E. coli ribosome operon, β-lactamase gene (bla), replication initiation site (ori), including unique recognition sites by restriction endonucleases with the following coordinates: EcoRI - 1, BamHI - 147, HindIII - 418, Pvu I - 1352. 2. An Escherichia coli cell transformed with recombinant plasmid DNA pHINS05 according to claim 1, is a producer of a hybrid polypeptide containing human proinsulin. 3. The bacterial strain Escherichia coli JM109 / pHINS05 is a producer of a hybrid polypeptide containing human proinsulin. (Collection of microorganisms of OJSC National Biotechnologies No. 05).