Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors - PubMed (original) (raw)

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

P Sheffield et al. Protein Expr Purif. 1999 Feb.

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.

Copyright 1999 Academic Press.

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