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Smith/Birnstiel mapping of genome rearrangements in Pseudomonas aeruginosa
Author(s) -
Heuer Thomas,
Bürger Christiane,
Tümmler Burkhard
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150190407
Subject(s) - biology , genome , locus (genetics) , restriction map , genetics , hypervariable region , restriction fragment , terminal restriction fragment length polymorphism , restriction enzyme , pseudomonas aeruginosa , restriction site , microbiology and biotechnology , restriction fragment length polymorphism , gene , plasmid , polymerase chain reaction , bacteria
A novel application of the Smith/Birnstiel technique is presented for the analysis of intraspecies genomic diversity in small genomes. Rare‐cutter total/ N frequent‐cutter partial restriction digestions are separated in N separate lanes by pulsed‐field gel electrophoresis, blotted and hybridized with rare‐cutter fragment ends as probes. The evaluation of the autoradiogram results in high‐resolution restriction maps of 10–200 kbp regions. The technique was applied to the analysis of genome rearrangements in Pseudomonas aeruginosa strains. Comparison of the region encoding the tryptophan biosynthesis genes in the PAO and the IATS serotype 5 strains revealed that shared sequence characterized by almost identical restriction fingerprints was interrupted in the serotype 5 strain by small islands displaying strain‐specific restriction site signatures. A multistep rearrangement in a hypervariable chromosome region downstream of the phn locus was detected in serial airway isolates from a patient with cystic fibrosis.