IngentaConnect Long-Term, Stable Expression of Green Fluorescent Protein in Mamm...

Authors: Gubin A.N.; Reddy B.; Njoroge J.M.; Miller J.L.

Source: Biochemical and Biophysical Research Communications, Volume 236, Number 2, July 1997 , pp. 347-350(4)

Abstract:

Despite the proven utility of green fluorescent protein (GFP) as a reporter molecule for transient gene expression, the adequacy of this marker for models requiring durable, high-level gene expression has not been fully tested. To address this issue, we performed the transfection of Chinese Hamster Ovary (CHO) cells with plasmid DNA encoding both GFP and neomycin phosphotransferase (neo) cassettes. The expression of GFP was measured after the cells were cultured in the presence or absence of G418-mediated selective pressure. After removal of G418 from the growth medium, the percentage of pooled G418 resistant transfectants which co-expressed the GFP transgene increased or remained unchanged. Flow cytometric and visual isolation of GFP-expressing cells was possible without continued selection in G418. One cloned cell line, C463, maintained high-level green fluorescence for 18 weeks in G418 and an additional 12 weeks in nonselective medium. Our data suggest expression of GFP does not confer a growth disadvantage in mammalian cells.

Language: English

Document Type: Research article

Affiliations: National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville Pike, Bethesda, Maryland, 20892:

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Long-Term, Stable Expression of Green Fluorescent Protein in Mammalian Cells