Placing the RPL32 Promoter Upstream of a Second Promoter Results in a Strongly Increased Number of Stably Transfected Mammalian Cell Lines That Display High Protein Expression Levels
| Autor: | John Antonius Verhees, Femke Hoeksema, Arie P. Otte, Karien M. Hamer, Michel Siep |
|---|---|
| Přispěvatelé: | Epigenetic Regulation of Gene Expression (inactive) (SILS, FNWI) |
| Rok vydání: | 2010 |
| Předmět: | |
| Zdroj: | Biotechnology Research International Biotechnology Research International, 2011 |
| ISSN: | 2090-3146 2090-3138 |
| Popis: | The use of high stringency selection systems commonly results in a strongly diminished number of stably transfected mammalian cell lines. Here we placed twelve different promoters upstream of an adjacent primary promoter and tested whether this might result in an increased number of colonies; this is in the context of a stringent selection system. We found that only the promoter of the human ribosomal protein, RPL32, induced a high number of colonies in CHO-DG44 cells. This phenomenon was observed when the RPL32 promoter was combined with the CMV, SV40, EF1-α, and theβ-actin promoters. In addition, these colonies displayed high protein expression levels. The RPL32 promoter had to be functionally intact, since the deletion of a small region upstream of the transcription start site demolished its positive action. We conclude that adding the RPL32 promoter to an expression cassettein cismay be a powerful tool to augment gene expression levels. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|