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Protein Expr Purif. 1999 Feb;15(1):34-9.

Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors.

Author information

1
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia, 22908, USA. pjs4n@virginia.edu

Abstract

We describe the construction of expression vectors based on three of the most frequently used gene fusion affinity tags [glutathione S-transferase (GST), maltose binding protein (MBP), and the His6 peptide]. The polylinkers of pGEX4T1, pMal-c2, and a pET vector were replaced with the polylinker isolated from the baculovirus expression plasmid pFastBac. Once appropriate restriction sites have been introduced into a gene, it can be fused to all three affinity tags with little effort, allowing expression-screening experiments to be performed efficiently. We discuss the development and use of these vectors with respect to overcoming purification problems encountered for the RhoA GDP/GTP nucleotide dissociation inhibitor (RhoGDI) and their advantages over commercially available expression vectors.

PMID:
10024467
DOI:
10.1006/prep.1998.1003
[Indexed for MEDLINE]

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