RT Journal Article SR Electronic(1) A1 Liao, Xiaowen A1 Charlebois, Isabelle A1 Ouellet, Catherine A1 Morency, Marie-Josée A1 Dewar, Ken A1 Lightfoot, Jeff A1 Foster, Jennifer A1 Siehnel, Richard A1 Schweizer, Herbert A1 Lam, Joseph S. A1 Hancock, Robert E. W. A1 Levesque, Roger C.YR 1996 T1 Physical mapping of 32 genetic markers on the Pseudomonas aeruginosa PAO1 chromosome JF Microbiology, VO 142 IS 1 SP 79 OP 86 DO https://doi.org/10.1099/13500872-142-1-79 PB Microbiology Society, SN 1465-2080, AB The Pseudomonas aeruginosa chromosome was fractionated with the enzymes Spel and Dpnl, 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., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/13500872-142-1-79