Bacterial comparative genomic hybridization: a method for directly identifying lateral gene transfer.
Horizontally transferred DNA is largely responsible for the dissemination of virulence traits amongst bacteria. Rapid identification of acquired DNA remains difficult as whole-genome sequencing of outbreak strains is impractical, and microarray-based approaches, while powerful, are limited to genes present only in the reference strains. Here we present a novel bacterial comparative genomic hybridization method that directly compares the genomes of related strains at sub-kilobase resolution in order to identify acquired DNA. Bacterial comparative genomic hybridization utilizes the concept of metaphase chromosome comparative genomic hybridization, and exploits the resolving power of two-dimensional DNA electrophoresis. Comparison of isogenic variants of the pathogen Pseudomonas aeruginosa detected a single-copy gene insertion responsible for gentamicin resistance.
Malloff
C A
CA
Department of Pathology, University of British Columbia, G-227-221 Westbrook Mall, Vancouver, British Columbia, V6T 2B5, Canada.
Fernandez
R C
RC
Lam
W L
WL
D001419:Bacteria / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D022761:Gene Transfer, Horizontal; D017403:In Situ Hybridization / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000235:genetics; D015183:Restriction Mapping
D004269:DNA, Bacterial; D015252:Deoxyribonucleases, Type II Site-Specific; C032229:GANTC-specific type II deoxyribonucleases