J. Bacteriol., Apr 1996, 1908-1913, Vol 178, No. 7
Copyright © 1996, American Society for Microbiology

Physical map of the genome of Planctomyces limnophilus, a representative of the phylogenetically distinct planctomycete lineage

N Ward-Rainey, FA Rainey, EM Wellington and E Stackebrandt
Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany.

A physical map of the chromosome of Planctomyces limnophilus DSM 3776T was constructed by pulsed-field gel electrophoresis techniques. A total of 32 cleavage sites for the rare-cutting restriction endonucleases PacI, PmeI, and SwaI were located on the chromosome, which was shown to be circular and approximately 5.2 Mbp in size. An extrachromosomal element was detected but was found not to be cleaved by any of the enzymes used in the analysis of the chromosome. The order of the fragments on the chromosome was determined by hybridization of excised, labelled restriction fragments to Southern blots of pulsed-field gel electrophoresis-separated restriction digests. Seven genetic markers, rrs, rrl, atpD, tuf, gyrB, rpoD, and dnaK, on the chromosome were located by hybridization. Probes for all genetic markers were obtained by PCR. For five of these markers, probes were constructed by PCR with degenerate primers targeting conserved sequences. The arrangement of the genetic markers was compared with that found in other bacteria.

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