RRC ID 33380
Author Sharrocks AD.
Title A T7 expression vector for producing N- and C-terminal fusion proteins with glutathione S-transferase.
Journal Gene
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.
Volume 138(1-2)
Pages 105-8
Published 1994-1-28
PII 0378-1119(94)90789-7
PMID 8125285
MeSH Amino Acid Sequence Animals Bacteriophage T7 / genetics* Base Sequence Cloning, Molecular / methods* Consensus Sequence DNA Primers DNA-Binding Proteins / biosynthesis* DNA-Binding Proteins / genetics Escherichia coli / genetics Escherichia coli / metabolism* Genetic Vectors* Glutathione Transferase / biosynthesis* MADS Domain Proteins MEF2 Transcription Factors Molecular Sequence Data Myogenic Regulatory Factors Nuclear Proteins / biosynthesis* Nuclear Proteins / genetics Plasmids Polymerase Chain Reaction Recombinant Fusion Proteins / biosynthesis* Restriction Mapping Schistosoma japonicum / enzymology Schistosoma japonicum / genetics* Thrombin / metabolism Transcription Factors / biosynthesis* Transcription Factors / genetics
IF 2.498
Times Cited 59
Prokaryotes E. coli pETGEXCT