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Title

Engineering Gram-Negative Microbial Cell Factories Using Transposon Vectors

AuthorsMartínez-García, Esteban; Aparicio, Tomás; Lorenzo, Víctor de; Nikel, Pablo I.
KeywordsMini-transposon
Tn5 transposon
Pseudomonas putida
Escherichia coli
Synthetic biology
Metabolic engineering
Microbial cell factory
Genome editing
Issue DateOct-2016
PublisherSpringer
CitationIn Vitro Mutagenesis : 273-293 (2016)
SeriesMethods in Molecular Biology
1498
AbstractThe 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.
Publisher version (URL)http://dx.doi.org/10.1007/978-1-4939-6472-7_18
URIhttp://hdl.handle.net/10261/146960
DOI10.1007/978-1-4939-6472-7_18
ISSN978-1-4939-6470-3
E-ISSN978-1-4939-6472-7
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