TY - JOUR
T1 - Physical mapping of 32 genetic markers on the Pseudomonas aeruginosa PAO1 chromosome
AU - Liao, Xiaowen
AU - Charlebois, Isabelle
AU - Ouellet, Catherine
AU - Morency, Marie Josée
AU - Dewar, Ken
AU - Lightfoot, Jeff
AU - Foster, Jennifer
AU - Siehnel, Richard
AU - Schweizer, Herbert
AU - Lam, Joseph S.
AU - Hancock, Robert E.W.
AU - Levesque, Roger C.
PY - 1996/1
Y1 - 1996/1
N2 - 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, pbpS, 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.
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, pbpS, 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.
KW - Genetic markers
KW - Physical mapping
KW - Pseudomonas aeruginosa chromosome
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U2 - 10.1099/13500872-142-1-79
DO - 10.1099/13500872-142-1-79
M3 - Article
C2 - 8581173
AN - SCOPUS:13344283427
SN - 1350-0872
VL - 142
SP - 79
EP - 86
JO - Microbiology
JF - Microbiology
IS - 1
ER -