title: Essential genes in the infection model of Pseudomonas aeruginosa-PCR-based signature-tagged mutagenesis.
abstract: PCR-based signature tagged mutagenesis is an "en masse" screening technique based upon unique oligonucleotide tags (molecular barcodes) for identification of genes that will diminish or enhance maintenance of an organism in a specific ecological niche or environment. PCR-based STM applied to Pseudomonas aeruginosa permitted the identification of genes essential for in vivo maintenance by transposon insertion and negative selection in a mixed population of bacterial mutants. The innovative adaptations and refinement of the technology presented here with P. aeruginosa STM mutants selected in the rat model of chronic lung infection have given critical information about genes essential for causing a chronic infection and a wealth of information about biological processes in vivo. The additional use of competitive index analysis for measurement of the level of virulence in vivo, microarray-based screening of selected prioritized STM mutants coupled to metabolomics analysis can now be attempted systematically on a genomic scale. PCR-based STM and combined whole-genome methods can also be applied to any organism having selectable phenotypes for screening.
authors: Institut de biologie intégrative et des systèmes (IBIS), Faculté de Médecine, Université Laval, Pavillon Charles-Eugène Marchand, 1030 Avenue de la médecine, Québec, QC, Canada, G1K 7P4.
authors: Irena
authors: I
authors: Kukavica-Ibrulj
authors: Roger C
authors: RC
authors: Levesque
mesh_terms: D000818:Animals; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D018274:Electroporation; D055786:Gene Knockout Techniques; D015723:Gene Library; D005798:Genes, Bacterial; D020043:Genes, Essential; D008297:Male; D016296:Mutagenesis / Q000235:genetics*; D009154:Mutation; D009841:Oligonucleotides / Q000378:metabolism*; D016366:Open Reading Frames / Q000235:genetics; D020161:Physical Chromosome Mapping; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D017207:Rats, Sprague-Dawley
chemical_list: D009841:Oligonucleotides