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Protein Expr Purif. 2008 Dec;62(2):185-9. doi: 10.1016/j.pep.2008.07.016. Epub 2008 Aug 8.

Expression and purification of the ligand-binding domain of peroxisome proliferator-activated receptor alpha (PPARalpha).

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  • 1Department of Pharmaceutical Chemistry & Bioanalytics, Institute of Pharmacy, Martin-Luther-Universität Halle-Wittenberg, Halle, Saale, Germany.

Abstract

We describe the expression and purification of the C-terminally His-tagged ligand-binding domain of the peroxisome proliferator-activated receptor alpha (PPARalpha-LBD). Using a modified expression and purification strategy a five times higher protein yield was achieved compared to existing protocols. In summary, a modified Escherichia coli strain was used, which allows rare codons to be expressed more efficiently, and moreover, conditions for cell growth and cell lysis were improved. Protein purification was achieved in a two-step approach using nickel affinity chromatography followed by anion exchange chromatography. The identity of PPARalpha-LBD was confirmed by online-nano-high performance liquid chromatography/matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (nano-HPLC/MALDI-TOF/TOF-MS).

PMID:
18755277
[PubMed - indexed for MEDLINE]
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