FIELD: genetic engineering, molecular biology, biochemistry.
SUBSTANCE: method involves construction of recombinant plasmid DNA pES4-4 with molecular mass 3.64 Mda (5917 base pairs) and consisting of Nde I/Bam HI-fragment of DNA comprising sequence of artificial gene of recombinant human interferon α-2b, β-lactamase gene and Nde I/Bam HI-fragment of plasmid pET22b(+)DNA comprising promoter and transcription terminator of T7-RNA polymerase, translation enhancer of phage T7 gene 10. Plasmid pES4-4 comprises β-lactamase as a genetic marker determining resistance of E. coli cells transformed with this plasmid to ampicillin, and unique recognition sites for endonuclease restriction enzymes located in the following distance to the right from site Nde I: Xba I - 38 base pairs; Hpa I - 1332 base pairs; Pst I - 4065 base pairs; Pvu I - 4190 base pairs, and Xho I - 5363 base pairs. Escherichia coli cells are transformed with prepared plasmid and the strain E. coli BDEES4 is obtained that is a super producer of recombinant human interferon α-2b. Invention provides preparing recombinant human interferon α-2b with high yield and by simplified technology. Invention can be used for preparing recombinant human interferon α-2b.
EFFECT: improved preparing method.
3 cl, 2 dwg, 2 ex
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