Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors
Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors
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DOI：
10.1006/prep.1998.1003
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发表时间：
1999-02-01
影响因子：
1.6
通讯作者：
Derewenda, Z
中科院分区：
文献类型：
--
作者：
Sheffield, P;Garrard, S;Derewenda, Z
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 His(6) 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, (C) 1999 Academic Press.
