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Nucleic Acids Research, 1983, Vol. 11, No. 3 687-706
© 1983

MOLECULAR BIOLOGY

Constitutive, long-term production of human interferons by hamster cells contalning multiple copies of a cloned interferon gene

Joel Haynes* and Charles Weissman

Institut for Molekularbiologie I, Universitai Zurich 8093,Zurich, Switzerland

Received December 7, 1982. Accepted January 4, 1983.

Hybrid plasmids containing the mouse dihydrofolate re-ductase (dhfr) and a human interferon (either IFN-{alpha}5 or IFN-{gamma}JO coding sequence under the control of viral promoters were trans-fected into dhfr– Chinese hamster ovary (CHO) cells. Dhfr+ colonies produced IFN at 10-1000 units-ml–1-day–1. Clones selected in methotrexate had a 20-50-fold increase in the IFN-a5 and dhfr DNA and mRNA content and secreted IFN at 20,000-100,000 units.ml–1.day–1"1. SDS-polyacrylamide gel electrophoresis of partially purified 35s-HuIFN-{gamma} from CHO cells showed a multiplet of labeled bands with a mobility corresponding to 22,400 to 23,400 daltons which was absent in the supernatants of non-transformed CHO cells. The higher apparent molecular weight of human IFN-J{gamma} from CHO cells as compared to that of human IFN-{gamma} from E.coli (about 18,800) suggests that the former was glycosylated.

*Present address: Connaught Research Institute, 1755 Steeles Avenue West, Willowdale, Ontario M2R 3T4, Canada


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