Characterisation of a DNA Polymerase Highly Mutated Along the Template Binding Interface
| Autor: | Jean-Luc Jestin, Sophie Vichier-Guerre |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Phage display
Thermus aquaticus biology DNA polymerase RNA-Directed DNA Polymerase Bioengineering DNA-Directed DNA Polymerase Templates Genetic Protein engineering biology.organism_classification Directed evolution Applied Microbiology and Biotechnology Biochemistry Molecular biology Reverse transcriptase Kinetics Mutation biology.protein Thermus Molecular Biology Polymerase Biotechnology |
| Zdroj: | Molecular Biotechnology. 46:58-62 |
| ISSN: | 1559-0305 1073-6085 |
| DOI: | 10.1007/s12033-010-9275-4 |
| Popis: | Phage display establishes a link between a polypeptide and its corresponding gene. It has been much used for the isolation of proteins binding to chosen molecular targets. A second link was designed more recently between a phage-displayed enzyme and its reaction product. Affinity chromatography for the product then allows the isolation of catalytically active enzymes and of their genes. Using this strategy, a polymerase with 15 mutations was selected by directed evolution of Thermus aquaticus DNA polymerase I. The kinetic characterisation reported here highlights the variant's broad template specificity and classifies this enzyme as a thermostable DNA-dependent and RNA-dependent DNA-polymerase. |
| Databáze: | OpenAIRE |
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