Prion protein expression in Chinese hamster ovary cells using a glutamine synthetase selection and amplification system
| dc.contributor.author | Blochberger, Thomas C. | |
| dc.contributor.author | Cooper, Carl | |
| dc.contributor.author | Peretz, David | |
| dc.contributor.author | Tatzelt, Jorg | |
| dc.date.accessioned | 2016-06-03T19:08:14Z | |
| dc.date.available | 2016-06-03T19:08:14Z | |
| dc.date.issued | 1997-09-05 | |
| dc.description | 10 pages | en_US |
| dc.description.abstract | Syrian hamster prion protein (PrPc) and a truncated Syrian hamster prion protein lacking the glycosylphosphatidylinositol (GPI) anchor C-terminal signal sequence (GPI~) were expressed in Chinese hamster ovary cells using a glutamine synthetase selection and amplification system. The CHO cell clones expressing the GPI" PrP secreted the majority of the protein into the media, whereas most of the PrP produced by clones expressing the full-length protein with the GPI anchor was located on the cell surface, as demonstrated by its release upon treatment with phosphatidylinositol-specific phospholipase C (PIPLC). A cell clone that expressed the highest levels of full length PrP was subcloned to obtain clone 30C3-1. PrP from clone 30C3-1 was shown to be sensitive to proteolysis by proteinase K and to react with monoclonal and polyclonal antibodies that recognize native PrPc. The recombinant PrP migrated as a diffuse band of 19-40 kDa but removal of the N-linked oligosaccharides with peptide N-glycosidase F (PNGase F) revealed three protein species of 19, 17 and 15 kDa. The 19 kDa band corresponding to deglycosylated full-length PrP was quantified and found to be expressed at a level - 14-fold higher than that of PrPc found in Syrian hamster brain. | en_US |
| dc.identifier.citation | Blochberger, T. C., Cooper, C., Peretz, D., Tatzlet, J., Griffith, O. H., Baldwin, M. A. and Prusiner, S. B. (1997). Prion protein expression in Chinese hamster ovary cells using a glutamine synthetase selection and amplification system, Protein Engineering, 10, 1465-1473. | en_US |
| dc.identifier.uri | https://hdl.handle.net/1794/19935 | |
| dc.language.iso | en_US | en_US |
| dc.publisher | Protein Engineering | en_US |
| dc.rights | Creative Commons BY-NC-ND 4.0-US | en_US |
| dc.title | Prion protein expression in Chinese hamster ovary cells using a glutamine synthetase selection and amplification system | en_US |
| dc.type | Article | en_US |