Chapter title |
Engineering Gram-Negative Microbial Cell Factories Using Transposon Vectors.
|
---|---|
Chapter number | 18 |
Book title |
In Vitro Mutagenesis
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6472-7_18 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6470-3, 978-1-4939-6472-7
|
Authors |
Esteban Martínez-García, Tomás Aparicio, Víctor de Lorenzo, Pablo I. Nikel |
Editors |
Andrew Reeves |
Abstract |
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. |
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