Expressed sequence tags (ESTs) from the Arabidopsis thaliana sequencing project were used to construct a genetic RFLP map for Brassica oleracea. Of the 110 A. thaliana ESTs tested, 95 were found to be informative RFLP probes in map construction. In total, 212 new loci corresponding to the 95 ESTs were added to the existing genetic map of B. oleracea. The enriched map covers all nine basic linkage groups and confirms that the chromosomes of B. oleracea and A. thaliana are similar in linear organization. However, varying levels of sequence conservation between the chromosomes of B. oleracea and A. thaliana were detected in different regions of the genomes. Long conserved regions encompassing entire chromosome arms in both genomes were identified; these are probably shared by descent. On the other hand, extensive rearrangements were observed in numerous chromosome regions, producing a mosaic of A. thaliana-like segments in the genome of Brassica. The presence of extensive chromosome duplication in A. thaliana was taken into consideration in the construction of the comparative maps of B. oleracea and A. thaliana.