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

Authors
  • Aubert, Daniel F
  • Hamad, Mohamad A
  • Valvano, Miguel A
Type
Published Article
Journal
Methods in Molecular Biology
Publication Date
Jan 01, 2014
Volume
1197
Pages
311–327
Identifiers
DOI: 10.1007/978-1-4939-1261-2_18
PMID: 25172289
Source
Medline
License
Unknown

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.

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