Definition of the zebrafish genome using flow cytometry and cytogenetic mapping

Autor: Zhou Yi, Rogers Jane, Humphray Sean, Scott Carol E, Zepeda Cinthya, Ng Bee, Chi Jianxiang, Maguire Sean F, Dallaire Stephanie, Banerjee Ruby, Adeniyi Adeola, Freeman Jennifer L, Zon Leonard I, Carter Nigel P, Yang Fengtang, Lee Charles
Jazyk: angličtina
Rok vydání: 2007
Předmět:
Zdroj: BMC Genomics, Vol 8, Iss 1, p 195 (2007)
Druh dokumentu: article
ISSN: 1471-2164
DOI: 10.1186/1471-2164-8-195
Popis: Abstract Background The zebrafish (Danio rerio) is an important vertebrate model organism system for biomedical research. The syntenic conservation between the zebrafish and human genome allows one to investigate the function of human genes using the zebrafish model. To facilitate analysis of the zebrafish genome, genetic maps have been constructed and sequence annotation of a reference zebrafish genome is ongoing. However, the duplicative nature of teleost genomes, including the zebrafish, complicates accurate assembly and annotation of a representative genome sequence. Cytogenetic approaches provide "anchors" that can be integrated with accumulating genomic data. Results Here, we cytogenetically define the zebrafish genome by first estimating the size of each linkage group (LG) chromosome using flow cytometry, followed by the cytogenetic mapping of 575 bacterial artificial chromosome (BAC) clones onto metaphase chromosomes. Of the 575 BAC clones, 544 clones localized to apparently unique chromosomal locations. 93.8% of these clones were assigned to a specific LG chromosome location using fluorescence in situ hybridization (FISH) and compared to the LG chromosome assignment reported in the zebrafish genome databases. Thirty-one BAC clones localized to multiple chromosomal locations in several different hybridization patterns. From these data, a refined second generation probe panel for each LG chromosome was also constructed. Conclusion The chromosomal mapping of the 575 large-insert DNA clones allows for these clones to be integrated into existing zebrafish mapping data. An accurately annotated zebrafish reference genome serves as a valuable resource for investigating the molecular basis of human diseases using zebrafish mutant models.
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