A markerless deletion method for genetic manipulation of Burkholderia cenocepacia and other multidrug-resistant gram-negative bacteria

Daniel F. Aubert, Mohamad A. Hamad, Miguel A. Valvano

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Genetic manipulation of multidrug-resistant bacteria is often difficult and hinders progress in understanding their physiology and pathogenesis. This book chapter highlights advances in genetic manipulation of Burkholderia cenocepacia, which are also applicable to other members of the Burkholderia cepacia complex and multidrug-resistant gram-negative bacteria of other genera. The method detailed here is based on the I-SceI homing endonuclease system, which can be efficiently used for chromosomal integration, deletion, and genetic replacement. This system creates markerless mutations and insertions without leaving a genetic scar and thus can be reused successively to generate multiple modifications in the same strain.

Original languageEnglish
Pages (from-to)311-327
Number of pages17
JournalMethods in Molecular Biology
Volume1197
DOIs
Publication statusPublished - 18 Jul 2014

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