title: A markerless deletion method for genetic manipulation of Burkholderia cenocepacia and other multidrug-resistant gram-negative bacteria.
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.
authors.affiliation: Department of Microbiology and Immunology, Centre for Human Immunology, University of Western Ontario, London, ON, Canada, N6A 5C1.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Aubert
authors.forename: Mohamad A
authors.initials: MA
authors.lastname: Hamad
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valvano
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D057508:Burkholderia cenocepacia / Q000187:drug effects / Q000235:genetics*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins