Nervous and Nonnervous Cell Transduction by Recombinant Adenoviruses That Inducibly Express the Human PrP
| Autor: | Franck Mouthon, Bernard Klonjkowski, Samuel Arrabal, Jean-Philippe Deslys, Dominique Dormont, Marc Eloit, Maryse Touchard |
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| Rok vydání: | 2001 |
| Předmět: |
Glycosylation
Prions animal diseases Genetic Vectors Biophysics CHO Cells In Vitro Techniques medicine.disease_cause Hippocampus Biochemistry Adenoviridae Cell Line Mice Transduction (genetics) chemistry.chemical_compound Genes Reporter Transduction Genetic Cerebellum Cricetinae medicine Animals Humans RNA Messenger Promoter Regions Genetic Molecular Biology Mice Knockout Neurons Regulation of gene expression biology Chinese hamster ovary cell Cell Membrane Cell Biology Tetracycline Flow Cytometry Proteinase K Molecular biology In vitro nervous system diseases Amino Acid Substitution Gene Expression Regulation chemistry Cell culture Doxycycline Mutagenesis Site-Directed biology.protein Plasmids |
| Zdroj: | Biochemical and Biophysical Research Communications. 285:623-632 |
| ISSN: | 0006-291X |
| Popis: | The study of the prion protein (PrP) physiological functions or its specific role in transmissible spongiform encephalopathies (TSE) requires new tools, particularly those able to induce PrP overexpression in a large range of cells, in vivo as well as in vitro. Here we describe the construction of two recombinant adenoviruses encoding the human PrP either with a valine at position 129 (AdTRVal) or a methionine (AdTRMet). Both genes were put under the control of the tetracycline-responsive promoter, allowing tight regulation of PrP expression. AdTRVal and AdTRMet induced high expression of the human PrP in CHO-KI cells and in organotypic brain slices in culture. The proteins expressed from these viruses exhibited a glycosylphosphatidyl inositol (GPI) anchor, proper glycosylation and sensitivity to proteinase K digestion. AdTRVal and AdTRMet will allow future studies on the human PrP and on the role of the codon 129 polyphormism in human TSE. |
| Databáze: | OpenAIRE |
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