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PMID: 6188107 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Nucleic acids research ·Vol. 11 ·No. 3 ·1983-02-11 ·Pages 687-706

Haynes J, Weissmann C

Abstract

Hybrid plasmids containing the mouse dihydrofolate reductase (dhfr) and a human interferon (either IFN-alpha 5 or IFN-gamma) coding sequence under the control of viral promoters were transfected into dhfr- Chinese hamster ovary (CHO) cells. dhfr+ colonies produced IFN at 10-1000 units X ml-1 X day-1. Clones selected in methotrexate had a 20-50-fold increase in the IFN-alpha 5 and dhfr DNA and mRNA content and secreted IFN at 20,000-100,000 units X ml-1 X day-1. SDS-polyacrylamide gel electrophoresis of partially purified 35S-HuIFN-gamma from CHO cells showed a multiple 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-gamma from CHO cells as compared to that of human IFN-gamma from E. coli (about 18,800) suggests that the former was glycosylated.

MeSH Terms
Animals Cell Line Cloning, Molecular Cricetinae Cricetulus DNA Restriction Enzymes Female Genes Humans Interferon Type I/genetics Interferon-gamma/genetics Interferons/isolation & purification Methionine/metabolism Molecular Weight Nucleic Acid Hybridization Ovary Plasmids RNA, Messenger/genetics Tetrahydrofolate Dehydrogenase/genetics
Chemicals
Interferon Type I RNA, Messenger Interferon-gamma Interferons Methionine Tetrahydrofolate Dehydrogenase DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Haynes J
Weissmann C
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.3.687
Published
1983-02-11
Pages
687-706
Language
English
Region
England
NLM ID
0411011
PMCID
PMC325746
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/11.3.687 PMC Full Text View journal details
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