Expression vector with two-step control by the cI-pR-Q-p'R-qut-t'R module of coliphage lambda
| Autor: | Vladimir V. Kravchenko, Irina P. Gileva, Lev A. Petrenko |
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| Rok vydání: | 1989 |
| Předmět: |
Expression vector
Transcription Genetic Genetic Vectors Temperature Repressor Promoter General Medicine Biology Lambda phage biology.organism_classification Molecular biology Bacteriophage lambda beta-Lactamases Gene product Gene Expression Regulation Lac Operon Transcription (biology) Gene expression Genetics Escherichia coli Cloning Molecular Promoter Regions Genetic Gene Plasmids |
| Zdroj: | Gene. 78(1) |
| ISSN: | 0378-1119 |
| Popis: | A plasmid expression vector (pCEQ3), using temperature-regulated transcription from the p ' R promoter of bacteriophage λ, has been constructed. The vector is derived from pBR327 in which the Eco RI- Cla I fragment of plasmid DNA is replaced with a 2.2-kb DNA module cI857- p ' R -Q- p ' R -qut- t ' R , consisting of two regions of the λ genome. The first region contains the repressor gene cl857 and promoter p ' R ; the second one contains gene Q and the late promoter p R . When the repressor protein, product of the cl857 gene, becomes temperatureinactivated, it allows the promoter p R to initiate the transcription of the Q gene. The product of the Q gene, in turn, acts as a positive regulator of transcription from promoter p ' R . The promoter activity of p R is fully repressed at a low temperature (30° C) and transcription from p ' R is terminated in the absence of Q gene product, but the shift of temperature up to 37°C is sufficient to make the transcription from the p R promoter highly active. Foreign genes can be inserted into the single Eco RI site downstream from the p ' R promoter. The resultant constructions express extremely high levels of the cloned gene product in Escherichia coli . |
| Databáze: | OpenAIRE |
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