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Wheat-germ cell-free production of prion proteins for solid-state NMR structural studies.
- Source :
-
New biotechnology [N Biotechnol] 2011 Apr 30; Vol. 28 (3), pp. 232-8. Date of Electronic Publication: 2010 Jul 04. - Publication Year :
- 2011
-
Abstract
- The expression of soluble, functional protein on a preparative scale poses a central challenge for structural studies. Cell-free protein expression has become a valuable alternative to cell-based methods, and allows today the expression of milligram quantities of protein. Its use is particularly attractive for NMR studies as it allows a multitude of isotopic labeling schemes. We have implemented and further developed protocols to prepare cell-free extracts from wheat germ to produce recombinant protein for solid-state NMR studies. Furthermore, we established the Renilla luciferase model to optimise and evaluate extract quality, and report first productions of the prions Ure2p and HET-s devoted to structural studies currently ongoing in our laboratories.<br /> (Copyright © 2010 Elsevier B.V. All rights reserved.)
- Subjects :
- Animals
Isotope Labeling
Luciferases, Renilla genetics
Luciferases, Renilla metabolism
Prions ultrastructure
Recombinant Proteins biosynthesis
Recombinant Proteins chemistry
Recombinant Proteins genetics
Cell-Free System metabolism
Nuclear Magnetic Resonance, Biomolecular methods
Prions chemistry
Prions metabolism
Triticum embryology
Subjects
Details
- Language :
- English
- ISSN :
- 1876-4347
- Volume :
- 28
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- New biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 20609396
- Full Text :
- https://doi.org/10.1016/j.nbt.2010.06.016