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Protein Expr Purif. 2011 Oct;79(2):258-62. doi: 10.1016/j.pep.2011.05.003. Epub 2011 May 14.

Production and single-step purification of EGFP and a biotinylated version of the Human Rhinovirus 14 3C protease.

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1
IBBS Biophysics Laboratories, School of Biological Sciences, University of Portsmouth, King Henry Building, King Henry I Street, Portsmouth PO1 2DY, Hampshire, United Kingdom. Jim.Youell@port.ac.uk

Abstract

The fluorescent reporter enhanced Green Fluorescent Protein (EGFP) has been used for assaying a wide range of biological activities ranging from gene expression, or localization of target proteins through to intermolecular interactions. However, over-production of this protein in Escherichia coli has resulted in the presence of inclusion bodies, which complicates recovery of the protein in significant quantities. In this paper, we describe a single-step method for isolating the protein from a Glutathione-S-Transferase (GST) fusion protein, release of the EGFP protein from the fusion was demonstrated using a biotinylated variant of Human Rhinovirus 14 3C protease that we have also constructed. We also suggest the potential uses of the biotinylated protease for bionanotechnology and synthetic biology.

PMID:
21605680
DOI:
10.1016/j.pep.2011.05.003
[Indexed for MEDLINE]
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