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 100 000 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.

Download full-text PDF

Source
http://dx.doi.org/10.1099/0022-1317-66-4-685DOI Listing

Publication Analysis

Top Keywords

murine interferon
8
interferon alpha
8
interferon
5
constitutive expression
4
expression murine
4
gene
4
alpha gene
4
gene hamster
4
hamster cells
4
cells characterization
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!