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Characterization of Japanese flounder karyotype by chromosome bandings and fluorescence in situ hybridization with DNA markers

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The chromosomes of Japanese flounder, Paralichthys olivaceus, were examined by conventional differential staining methods including G-, Q-, C-, silver (Ag)-, fluorochrome, and replication R-bandings and by fluorescence in situ hybridization (FISH) with 5S and 18S rDNAs and telomeric DNA as probes. Replication R-banding substantially made it possible to identify 24 homologous pairs by their RBG-banding pattern and relative length. Both rDNA loci were mapped to chromosome 1, where 5S and 18S rDNA loci were located at the centromeric region and secondary constriction, respectively. C-banding revealed that both rDNA loci were heterochromatic, and 18S rDNA loci were positive for chromomycin A3 but negative for 4′,6-diamidino-2-phenylindole (DAPI) staining. Telomeric FISH signals were observed at all chromosome ends and at the interstitial region of some chromosomes. The observed results were discussed in relation to the karyotype evolution in the order Pleuronectiformes.

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fluorescence in situ hybridization


silver-stained nucleolar organizing region(s)


propidium iodide



CMA3 :

chromomycin A3


polymerase chain reaction




R-bands by BrdU using Giemsa


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We thank Drs. H. Furuita and T. Kamaishi, National Research Institute of Aquaculture (NRIA), for kindly providing fish used. This study was supported in part by Grants-in-Aid from the Fishery Agency, and from the 21st Century COE Program from the Ministry of Education, Sports, Culture, Science and Technology, Japan.

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Correspondence to Atushi Fujiwara.

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Fujiwara, A., Fujiwara, M., Nishida-Umehara, C. et al. Characterization of Japanese flounder karyotype by chromosome bandings and fluorescence in situ hybridization with DNA markers. Genetica 131, 267–274 (2007).

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