FIELD: biotechnology, genetic engineering, molecular biology.
SUBSTANCE: invention can be used for preparing recombinant human growth hormone. Recombinant plasmid DNA pES1-6 encoding polypeptide with somatotropin amino acid sequence with molecular mass 3.66 MDa (5949 pair bases) consists of: Nde I/Xho I DNA fragment of plasmid pET22b(+) comprising transcription promoter and terminator of T7 RNA-polymerase, translation enhancer of phage T7 gene 10, β-lactamase gene and Nde I/Xho I DNA fragment comprising the sequence of artificial gene for recombinant somatotropin comprising: as genetic marker β-lactamase gene determining resistance of E. coli cells transformed with plasmid pSE1-6 to ampicillin; unique recognition sites for site-specific endonuclease restriction enzymes located at the following distance to the right of site Nde I : Xba I - 38 pair bases, Hpa I - 1332 pair bases, Pst I - 4065 pair bases, Pvu I - 4190 pair bases, Xho I - 5363 pair bases. Invention provides preparing the strain Escherichia coli BL21(DE3)/pES1-6 comprising recombinant plasmid DNA pES1-6 as a producer of recombinant somatotropin. Invention provides preparing recombinant somatotropin with high yield by the simplified technology.
EFFECT: improved preparing method, valuable biological properties of plasmid DNA.
3 cl, 2 dwg, 2 ex
