Evaluación de la actividad recombinasa e integrase sitio-específica de relaxasas conjugativas en bacterias y en células humanas
ABSTRACT: With this work we have got insight into the molecular characterization of site-specific recombination and integration reactions mediated by relaxases. We have carried out an extensive mutagenesis of trwC that suggest that the improvement of relaxase TrwC as recombinase by altering one or t...
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| Tipo de recurso: | tesis doctoral |
| Fecha de publicación: | 2015 |
| País: | España |
| Institución: | Universidad de Cantabria (UC) |
| Repositorio: | UCrea Repositorio Abierto de la Universidad de Cantabria |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.unican.es:10902/14008 |
| Acceso en línea: | http://hdl.handle.net/10902/14008 |
| Access Level: | acceso abierto |
| Palabra clave: | Conjugative relaxases Site-specific recombination Site-specific integration Genomic engineering Relaxasas conjugativas Recombinación sitio-específica Integración sitio-específica Ingeniería genética |
| Sumario: | ABSTRACT: With this work we have got insight into the molecular characterization of site-specific recombination and integration reactions mediated by relaxases. We have carried out an extensive mutagenesis of trwC that suggest that the improvement of relaxase TrwC as recombinase by altering one or two residues is not possible. Moreover, we have analysed the DNA requirements of the integration reaction by TrwC. Apart from TrwC, we have found that TraI proteins from plasmids F and pKM101 also have oriT-specific recombinase activity. In contrast, we did not detect F_TraI-mediated integration, suggesting a difference with TrwC. The potential use of TrwC for genomic engineering has been studied in human cells. We found that TrwC is active in human cells and it can act as a site-specific integrase, although not efficiently. The rate of integration of TrwC-driven DNA suggests that TrwC may be stabilizing the DNA in the human cell, favouring its random integration. |
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