Process for the production of unfused IGF-II protein in E. coli
AssigneeCiba-Geigy Corporation
InventorsAlbert Schmitz, Walter Marki
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Abstract
A process for the preparation of a recombinant IGF-II (rIGF-II) without a covalently attached foreign protein moiety and without N-terminal attached methionine or a derivative of methionine or of a salt of said IGF-II, rIGF-II produced by said method, hybrid vectors comprising DNA encoding said rIGF-II, hosts transformed with said vectors, and a process for the isolation of said rIGF-II from the host cell and refolding it into a biologically active form.
Claims15
A process for the production of a recombinant IGF-II without a covalently attached foreign protein moiety and without N-terminal attached methionine or a derivative of methionine, said process comprising:
- a) transforming a suitable strain of E. coli, said strain being a lon.sup.- and htpR.sup.- double mutant, with a hybrid vector comprising an expression cassette consisting of the following elements in the 5' to 3' direction, said elements which are operably linked:
- an inducible promoter, a ribosomal binding site, and the codon for the amino acid methionine linked in proper reading frame to a DNA sequence encoding a human IGF-II having the amino acid sequence of SEQ ID NO. 1,
- b) culturing said transformed strain under conditions suitable for production of a recombinant IGF-II from said expression cassette.
A hybrid vector comprising an expression cassette consisting of the following elements in the 5' to 3' direction, said elements which are operably linked:
- an inducible promoter, a ribosomal binding site, and the codon for the amino acid methionine linked in the proper reading frame to a DNA sequence encoding a human IGF-II having the amino acid sequence of Seq. ID No. 1.
Dependencies are inferred from the claim text.