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Nucleic Acids Research, 2000, Vol. 28, No. 10 2026-2033
© 2000 Oxford University Press

Encephalitozoon cuniculi (Microspora) genome: physical map and evidence for telomere-associated rDNA units on all chromosomes

Jean-François Brugère, Emmanuel Cornillot*, Guy Méténier, Aaron Bensimon1 and Christian P. Vivarès

Equipe de Parasitologie Moléculaire et Cellulaire, UPRES A CNRS 6023, Université Blaise Pascal, 63177 Aubière cedex, France and 1Laboratoire de Biophysique de l’ADN, Institut Pasteur, 75015 Paris, France

A restriction map of the 2.8-Mb genome of the unicellular eukaryote Encephalitozoon cuniculi (phylum Microspora), a mammal-infecting intracellular parasite, has been constructed using two restriction enzymes with 6 bp recognition sites (BssHII and MluI). The fragments resulting from either single digestions of the whole molecular karyotype or double digestions of 11 individual chromosomes have been separated by two-dimensional pulsed field gel electrophoresis (2D-PFGE) procedures. The average distance between successive restriction sites is ~19 kb. The terminal regions of the chromosomes show a common pattern covering ~15 kb and including one 16S–23S rDNA unit. Results of hybridisation and molecular combing experiments indicate a palindromic-like orientation of the two subtelomeric rDNA copies on each chromosome. We have also located 67 DNA markers (clones from a partial E.cuniculi genomic library) by hybridisation to restriction fragments. Partial or complete sequencing has revealed homologies with known protein-coding genes for 32 of these clones. Evidence for two homologous chromosomes III, with a size difference (3 kb) related to a subtelomeric deletion/insertion event, argues for diploidy of E.cuniculi. The physical map should be useful for both the whole genome sequencing project and studies on genome plasticity of this widespread parasite.

* To whom correspondence should be addressed. Tel: +33 4 73 40 74 71; Fax: +33 4 73 40 74 55; Email: emmanuel.cornillot@lbp.univ-bpclermont.fr


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