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J Gen Virol 66 (1985), 685-691; DOI 10.1099/0022-1317-66-4-685
© 1985 Society for General Microbiology
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Constitutive Expression of a Murine Interferon Alpha Gene in Hamster Cells and Characterization of Its Protein Product

Ellen C. Zwarthoff, I. Jaap Bosveld, Wim P. Vonk and Jan Trapman

Department of Pathology, Erasmus University, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands

The coding part of a murine interferon alpha (MuIFN-{alpha}) gene was cloned into an expression plasmid containing the simian virus 40 early promoter and the rabbit beta-globin polyadenylation signal. This construct was transfected into Chinese hamster ovary cells, together with a plasmid containing the Ecogpt gene as a selection marker. Resulting colonies were assayed for constitutive interferon production and analysed for integration of MuIFN-{alpha} genes. There was no obvious correlation between the number of genes integrated and the amount of interferon produced. The highest producer, designated CHO-pSV10EF-3, contained four copies of the mouse gene and constitutively secreted up to 100000 International Units of interferon per ml per day. The MuIFN-{alpha} subspecies produced by this clone was characterized by analysis of its antiviral activity on heterologous cells, heparin-Sepharose affinity chromatography and chromatofocusing. The results obtained indicate that it is identical or closely related to a minor component present in conventional MuIFN-{alpha} preparations.

Keywords: interferon (MuIFN-{alpha}) subspecies, CHO cells, constitutive expression

Received 23 August 1984; accepted 17 December 1984.




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