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Random Transposon Mutagenesis of Vibrio cholerae

  • Jyl S. MatsonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1839)

Abstract

Transposon-based random mutagenesis of bacterial genomes has proven to be a powerful genetic tool for the identification of genes and regulatory elements that contribute to specific phenotypes. One such approach that has been used in Vibrio cholerae for many years is the introduction of mariner transposons to generate random libraries of mutants. These libraries have been successfully used for a wide variety of genetic screens and selections in this important bacterial pathogen. Here we present a detailed protocol for the use of plasmid pFD1 (containing the mariner transposon magellan3) to create mutant libraries in V. cholerae.

Key words

Random mutagenesis Transposon mutagenesis Random-primed sequencing Nested PCR Mariner transposon 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Medical Microbiology and ImmunologyUniversity of Toledo College of Medicine and Life SciencesToledoUSA

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