Controlled high-level expression of the lon gene of Escherichia coli allows overproduction of Lon protease
| Autor: | P. M. Cullis, J Modha, A J Rivett, T M Razzaq, C D Thomas |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
DNA
Bacterial Protease La medicine.medical_treatment Molecular Sequence Data Biology medicine.disease_cause Gene product Plasmid ATP-Dependent Proteases Escherichia coli Genetics medicine Amino Acid Sequence Cloning Molecular Overproduction Gene Heat-Shock Proteins chemistry.chemical_classification Protease Base Sequence Escherichia coli Proteins Serine Endopeptidases Nucleic acid sequence General Medicine Molecular biology Enzyme Biochemistry chemistry bacteria Plasmids |
| Zdroj: | Gene. 136:237-242 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/0378-1119(93)90471-e |
| Popis: | Lon protease from Escherichia coli is an ATP-dependent protease which plays important roles in regulating the levels of specific proteins and in eliminating abnormal proteins. A major problem of working with Lon protease, the inability to substantially overproduce the enzyme, has been overcome by placing the lon gene under the control of an inducible trp promoter within a copy-number-controllable plasmid. Induction resulted in higher levels of production of the protease (approximately 100 micrograms/ml of cell culture) than were previously possible. The enzyme has been purified to apparent homogeneity and shown to possess the characteristic ATP-dependent proteolytic activity. Sequence verification during DNA manipulations revealed differences from two previously published sequences for the lon gene. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|