Engineering Gram-Negative Microbial Cell Factories Using Transposon Vectors

The construction of microbial cell factories à la carte largely depends on specialized molecular biology and synthetic biology tools needed to reprogram bacteria for modifying their existing functions or for bestowing them with new-to-Nature tasks. In this chapter, we document the use of a series of...

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Detalles Bibliográficos
Autores: Martínez-García, Esteban, Aparicio, Tomás, Lorenzo, Víctor de, Nikel, Pablo I.
Tipo de recurso: otro
Estado:Versión enviada para evaluación y publicación
Fecha de publicación:2016
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/146960
Acceso en línea:http://hdl.handle.net/10261/146960
Access Level:acceso abierto
Palabra clave:Mini-transposon
Tn5 transposon
Pseudomonas putida
Escherichia coli
Synthetic biology
Metabolic engineering
Microbial cell factory
Genome editing
Descripción
Sumario:The construction of microbial cell factories à la carte largely depends on specialized molecular biology and synthetic biology tools needed to reprogram bacteria for modifying their existing functions or for bestowing them with new-to-Nature tasks. In this chapter, we document the use of a series of broad-host-range mini-Tn5 vectors for the delivery of gene(s) into the chromosome of Gram-negative bacteria and for the generation of saturated, random mutagenesis libraries for studies of gene function. The application of these tailored mini-transposon vectors, which could also be used for chromosomal engineering of a wide variety of Gram-negative microorganisms, is demonstrated in the platform environmental bacterium Pseudomonas putida KT2440.