論文 - 詳細
RRC ID | 33380 |
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著者 | Sharrocks AD. |
タイトル | A T7 expression vector for producing N- and C-terminal fusion proteins with glutathione S-transferase. |
ジャーナル | 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. |
巻・号 | 138(1-2) |
ページ | 105-8 |
公開日 | 1994-1-28 |
DOI | 10.1016/0378-1119(94)90789-7 |
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 | 2.984 |
引用数 | 65 |
WOS 分野 | GENETICS & HEREDITY |
リソース情報 | |
原核生物(大腸菌) | pETGEXCT |