title: Physical mapping of 32 genetic markers on the Pseudomonas aeruginosa PAO1 chromosome.
abstract: The Pseudomonas aeruginosa chromosome was fractionated with the enzymes SpeI and DpnI, and genomic fragments were separated by PFGE and used for mapping a collection of 40 genes. This permitted the localization of 8 genes previously mapped and of 32 genes which had not been mapped. We showed that a careful search of databases and identification of sequences that were homologous to known genes could be used to design and synthesize DNA probes for the mapping of P. aeruginosa homologues by Southern hybridization with genomic fragments, resulting in definition of the locations of the aro-2, dapB, envA, mexA, groEL, oprH, oprM, oprP, ponA, rpoB and rpoH genetic markers. In addition, a combination of distinct DNA sources were utilized as radioactively labelled probes, including specific restriction fragments of the cloned genes (glpD, opdE, oprH, oprO, oprP, phoS), DNA fragments prepared by PCR, and single-stranded DNA prepared from phagemid libraries that had been randomly sequenced. We used a PCR approach to clone fragments of the putative yhhF, sucC, sucD, cypH, pbpB, murE, pbpC, soxR, ftsA, ftsZ and envA genes. Random sequencing of P. aeruginosa DNA from phagemid libraries and database searching permitted the cloning of sequences from the acoA, catR, hemD, pheS, proS, oprD, pyo and rpsB gene homologues. The described genomic methods permit the rapid mapping of the P. aeruginosa genome without linkage analysis.
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, 300-6174 University Boulevard, Vancouver BC, Canada V6T 1Z3.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Liao
authors.affiliation: Microbiologie Moléculaire et Génie des Protéines, Département de Microbiologie, Faculté de Médecine, Pavillon Charles-Eugène-Marchand, Université Laval, Ste-Foy, Québec, Canada G1K 7P4.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Charlebois
authors.affiliation: Microbiologie Moléculaire et Génie des Protéines, Département de Microbiologie, Faculté de Médecine, Pavillon Charles-Eugène-Marchand, Université Laval, Ste-Foy, Québec, Canada G1K 7P4.
authors.forename: Catherine
authors.initials: C
authors.lastname: Ouellet
authors.affiliation: Microbiologie Moléculaire et Génie des Protéines, Département de Microbiologie, Faculté de Médecine, Pavillon Charles-Eugène-Marchand, Université Laval, Ste-Foy, Québec, Canada G1K 7P4.
authors.forename: Marie-Josée
authors.initials: MJ
authors.lastname: Morency
authors.affiliation: Microbiologie Moléculaire et Génie des Protéines, Département de Microbiologie, Faculté de Médecine, Pavillon Charles-Eugène-Marchand, Université Laval, Ste-Foy, Québec, Canada G1K 7P4.
authors.forename: Ken
authors.initials: K
authors.lastname: Dewar
authors.affiliation: Microbiologie Moléculaire et Génie des Protéines, Département de Microbiologie, Faculté de Médecine, Pavillon Charles-Eugène-Marchand, Université Laval, Ste-Foy, Québec, Canada G1K 7P4.
authors.forename: Jeff
authors.initials: J
authors.lastname: Lightfoot
authors.affiliation: Department of Microbiology, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Foster
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, 300-6174 University Boulevard, Vancouver BC, Canada V6T 1Z3.
authors.forename: Richard
authors.initials: R
authors.lastname: Siehnel
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada T2N 4N1.
authors.forename: Herbert
authors.initials: H
authors.lastname: Schweizer
authors.affiliation: Department of Microbiology, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Lam
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, 300-6174 University Boulevard, Vancouver BC, Canada V6T 1Z3.
authors.forename: Robert E W
authors.initials: REW
authors.lastname: Hancock
authors.affiliation: Microbiologie Moléculaire et Génie des Protéines, Département de Microbiologie, Faculté de Médecine, Pavillon Charles-Eugène-Marchand, Université Laval, Ste-Foy, Québec, Canada G1K 7P4.
authors.forename: Roger C
authors.initials: RC
authors.lastname: Levesque
mesh_terms: D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D015870:Gene Expression; D005798:Genes, Bacterial; D005819:Genetic Markers; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D018076:DNA, Complementary; D005819:Genetic Markers; D015345:Oligonucleotide Probes; C020102:endodeoxyribonuclease DpnI; C056436:endodeoxyribonuclease SpeI; D015252:Deoxyribonucleases, Type II Site-Specific