FIELD: biotechnology.
SUBSTANCE: invention relates to biotechnology, particularly to genetic engineering, and is an in vitro constructed recombinant plasmid DNA containing two copies of a human granulocyte-macrophage colony-stimulating factor (GM-CSF) synthetic gene, Escherichia coli strain SG20050/p280_2GM is a producer of said polypeptide and a method of producing a polypeptide with GM-CSF properties. Recombinant plasmid DNA p280_2GM codes a polypeptide with properties of granulocyte-macrophage colony-stimulating factor (GM-CSF) of a person characterized by the following features: codes the amino acid sequence of the mature human GM-CSF; has molecular weight of 2.95 MDa (4,422 base pairs); consists of: PsTI/BamHI(pol)-fragment of plasmid DNA p280GM (1,484 base pairs) containing C-terminal part of beta-lactamase gene, tandem of promoters of tryptophan operon E. coli and synthetic gene GM-CSF of human, as well as EcoRI(pol)/PstI (2,938 base pairs) – a fragment of the same plasmid containing the synthetic human GM-CSF gene and the N-terminal portion of the beta-lactamase gene; contains: – tandem promoters of tryptophan operon E. coli; two copies of human synthetic GM-CSF gene; transcription terminator t0 of lambda phage; as a genetic marker, a beta-lactamase gene determining the resistance of E. coli transformed plasmid p280_2GM to ampicillin antibiotics; unique recognition sites by restriction endonucleases located at the following distances to the right from EcoRI site (279 base pairs) – BamHI – 912 nucleotides; PstI – 3,396 nucleotides.
EFFECT: invention increases output of polypeptide with properties of granulocyte-macrophage colony-stimulating factor.
3 cl, 8 dwg, 4 tbl, 15 ex
