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Title: Targeted mutagenesis of Burkholderia thailandensis and Burkholderia pseudomallei through natural transformation of PCR fragments
Authors: Metawee Thongdee
Larry A. Gallagher
Mark Schell
Tararaj Dharakul
Sirirurg Songsivilai
Colin Manoil
Mahidol University
University of Washington, Seattle
The University of Georgia
Keywords: Biochemistry, Genetics and Molecular Biology;Environmental Science;Immunology and Microbiology
Issue Date: 1-May-2008
Citation: Applied and Environmental Microbiology. Vol.74, No.10 (2008), 2985-2989
Abstract: Burkholderia pseudomallei is the causative agent of melioidosis, an overwhelming, rapidly fatal septic infection, and B. thailandensis is a closely related, less virulent species. Both organisms are naturally competent for DNA transformation, and this report describes a procedure exploiting this property for the rapid generation of marked deletion mutations by using PCR products. The method was employed to create 61 mutant strains. Several selectable elements were employed, including elements carrying loxP and FRT recombinase recognition sites to facilitate resistance marker excision. Chromosomal mutations could also be transferred readily between strains by transformation. The availability of simple procedures for creating defined chromosomal mutations and moving them between strains should facilitate genetic analysis of virulence and other traits of these two Burkholderia species. Copyright © 2008, American Society for Microbiology. All Rights Reserved.
ISSN: 00992240
Appears in Collections:Scopus 2006-2010

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