Engineering Gram-negative microbial cell factories using transposon vectors

Esteban Martínez-García, Tomás Aparicio, Victor de Lorenzo, Pablo Ivan Nikel

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review


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.
Original languageEnglish
Title of host publicationIn Vitro Mutagenesis
PublisherSpringer New York
Publication date2016
ISBN (Print)978-1-4939-6470-3
ISBN (Electronic)978-1-4939-6472-7
Publication statusPublished - 2016
Externally publishedYes
SeriesMethods in Molecular Biology


  • Mini-transposon
  • Tn 5 transposon
  • Pseudomonas putida
  • Escherichia coli
  • Synthetic biology
  • Metabolic engineering
  • Microbial cell factory
  • Genome editing


Dive into the research topics of 'Engineering Gram-negative microbial cell factories using transposon vectors'. Together they form a unique fingerprint.

Cite this