title: Screening and identification of potential sex-associated sequences in Danio rerio.
abstract: Current knowledge on zebrafish (Danio rerio) suggests that sex determination has a polygenic genetic basis in this species, although environmental factors may also be involved. This study aimed to identify sex-associated genomic regions using two different marker systems: inter-simple sequence repeats (ISSRs) and random-amplified polymorphic DNA (RAPDs). Two bulks were constructed: one with DNA from zebrafish females and the other from males; then, a total of 100 ISSR and 280 RAPD primers were tested. Three DNA fragments presenting sexual dimorphism (female-linked: OPA17436 and OPQ191027 ; male-linked: OPQ19951 ) were determined from sequential analysis of the bulks followed by assessment in individuals. These fragments were cloned and convert into the following sequenced characterized amplified regions (SCAR): DrSM_F1, DrSM_F2, and DrSM_M, which share identities with sequences located in chromosomes 2, 3, and 11 (Zv9), respectively. Using these potential markers in zebrafish samples it was possible to correctly identify 80% of the males (DrSM_M) and 100% of the females (DrSM_F1 + DrSM_F2) in the analyzed population.
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences, CITAB, University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Luzio
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences, CITAB, University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Coimbra
authors.affiliation: Departamento de Genética, Facultad de Biologia, Universidad Complutense, Madrid, Spain.
authors.forename: César
authors.initials: C
authors.lastname: Benito
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences, CITAB, University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
authors.forename: António A
authors.initials: AA
authors.lastname: Fontaínhas-Fernandes
authors.affiliation: Departamento de Genética e Biotecnologia (DGB), Escola de Ciências da Vida e do Ambiente (ECVA), Universidade de Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal.
authors.forename: Manuela
authors.initials: M
authors.lastname: Matos
mesh_terms: D000818:Animals; D017931:DNA Primers; D005260:Female; D008297:Male; D018895:Microsatellite Repeats; D019105:Random Amplified Polymorphic DNA Technique; D012727:Sex Characteristics; D019849:Sex Determination Processes / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics*
chemical_list: D017931:DNA Primers