Induction of recombinant gene expression in Escherichia coli using an alkaline pH shift
| Autor: | Kurt Poindexter, Richard B. Gayle |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
DNA
Recombinant Repressor medicine.disease_cause Cleavage (embryo) law.invention chemistry.chemical_compound law Gene expression Escherichia coli Genetics Protein biosynthesis medicine Promoter Regions Genetic Growth medium biology Temperature Gene Expression Regulation Bacterial General Medicine Hydrogen-Ion Concentration Lambda phage biology.organism_classification Molecular biology Rec A Recombinases Biochemistry chemistry Recombinant DNA |
| Zdroj: | Gene. 97:125-130 |
| ISSN: | 0378-1119 |
| Popis: | A commonly used approach to control recombinant protein production in Escherichia coli utilizes the lambda pL promoter-operator and the lambda repressor. Inactivation of the lambda repressor function allows transcription to proceed. However, induction of the RecA-mediated cleavage of lambda repressor by the addition of nalidixic acid or inactivation of a temperature-sensitive lambda repressor by growth at the nonpermissive temperature can have detrimental effects on protein production. This paper describes the use of an alkaline shift in the pH of the growth medium that allows expression of genes from the pL promoter in a RecA-independent manner. This procedure results in high-level production of recombinant protein. The pH shift is performed in the stationary phase of cell growth, using culture volumes ranging from 1-1000 ml. This method can result in the production of over 15-fold more active protein than when using a temperature shift. |
| Databáze: | OpenAIRE |
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