Elsevier

Gene

Volume 138, Issues 1–2, 28 January 1994, Pages 105-108
Gene

Short communication
A T7 expression vector for producing N- and C-terminal fusion proteins with glutathione S-transferase

https://doi.org/10.1016/0378-1119(94)90789-7Get rights and content

Abstract

The pGEX system for protein production in E. coli is widely used in molecular biology. A bacterial expression vector, pETGEXCT, which incorporates features of the pGEX and pET expression systems was designed. pETGEXCT allows the production of N- and C-terminal fusions to glutathione S-transferase (GST) under the tight control of the T7 promoter. Use of this vector can circumvent problems associated with unstable or inactive fusions to the N terminus of GST. Indeed, it is demonstrated that fusions to the N terminus of the eukaryotic DNA-binding protein, RSRFC4, cannot be tolerated. Fusion of RSRFC4 to the N terminus of GST in the pETGEXCT vector is a prerequisite to purify the RSRFC4 DNA-binding domain in an active form using glutathione-agarose affinity chromatography.

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