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Nucleic Acids Research, 1983, Vol. 11, No. 18 6419-6435
© 1983

MOLECULAR BIOLOGY

Chemical synthesis of a human interferon-{alpha}2, gene and its expression in Escherichia coli

M.D. Edge, A.R. Greene, G.R. Heathcliffe, V. E. Moore, N.J. Faulkner, R. Camble, N.N. Petter, P. Trueman, W. Schuch*, J. Hennam*, T.C. Atkinson, C.R. Newton and A.F. Markham

ICI Pharmaceuticals Division Alderley Park, Macclesfield, Cheshire, SK10 4TG *ICI Corporate Biosciences Group The Heath, Runcorn, Cheshire, WA7 4QE, UK

Received June 7, 1983. Revised August 16, 1983. Accepted August 16, 1983.

A 511-base pair DNA fragment encoding human interferon-a2 has been chemically synthesised and expressed from a lac UV5 and a synthetic trp promoter in Escherichia coli. The synthesis involved preparation of 68 ol1gode oxyr1bonuc1eotides and their enzymic ligation. The product expressed from the trp promoter system had high antiviralactivity and displayed biological effects similar to those of Namalwa Interferon on natural killer cellactivity and in a Daudi cell growth Inhibition assay. E.coli minicelis containing plasmid DNA with the synthetic IFN-{alpha}2 gene under trp promoter control produce a protein with the same electrophoretic mobility as a sample of authentic IFN-{alpha}2. The protein from E.coli cross-reacts with the monoclonal antibody NK-2 and was readily purified, close to homogeneity, by immunoadsorpt1on chromatography on NK-2 sepharose.


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