Percorrer por autor "Geisler, Robert"
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- Comparative analysis and characterization of expressed sequence tags in gilthead sea bream (Sparus aurata) liver and embryosPublication . Sarropoulou, Elena; Power, Deborah; Magoulas, Antonio; Geisler, Robert; Kotoulas, GeorgiosThe gilthead sea bream (Sparus aurata) is one of the main European aquaculture products and a prospective model species for the Sparidae, which includes several other commercially important species. Future selective breeding of aquaculture stocks will be heavily underpinned by molecular genetic techniques, especially by marker-assisted selection (MAS). Gene marker resources in marine fish species, however, lag behind those of other agricultural animals, and only scanty information exists about the genetic source of phenotypic variation and the identity of quantitative trait loci (QTL). In order to develop molecular resources in gilthead sea bream, complementary DNA libraries were constructed from liver and mixed embryo and larval stages by unidirectional cloning. A long-read expressed sequence tag (EST) database was generated, containing 1394 cDNA clones representing 852 unique cDNA sequence-reads. Tissue-specific patterns of gene expression were determined when grouped using the proposal for characterizing cellular component put forward by the Gene Ontology consortium. Transcripts encoding cytoskeletal proteins were most abundant in the embryonic/larval library, while the most abundant transcripts in the liver library encoded secreted and extracellular proteins. Of both libraries, 505 clones were sequenced in both orientations (5Vand 3Vend sequencing), where 226 clones were determined as full-length sequence reads. Cluster analysis of 3Vend-sequenced clones from both libraries revealed that alternative polyadenylation signals were utilized, although no evidence of alternative splicing was found. We report for the first time for gilthead sea bream or any sparid a transcriptional analysis of two tissues and briefly consider the utility of ESTs for characterizing tissue-specific expression profiles. D 2004 Elsevier B.V. All rights reserved.
- Gene expression profiling of gilthead sea bream during early development and detection of stress-related genes by the application of cDNA microarray technologyPublication . Sarropoulou, Elena; Kotoulas, Georgios; Power, Deborah; Geisler, RobertGene expression profiling of gilthead sea bream during early development and detection of stress-related genes by the application of cDNA microarray technology. Physiol Genomics 23: 182–191, 2005. First published July 26, 2005; doi:10.1152/physiolgenomics.00139.2005.—Large-scale gene expression studies were performed for one of the main European aquaculture species, the gilthead sea bream Sparus auratus L. For this purpose, a cDNA microarray containing 10,176 clones from a cDNA library of mixed embryonic and larval stages was constructed. In addition to its importance for aquaculture, the taxonomic position and the relatively small genome size of sea bream makes it a prospective model for evolutionary biology and comparative genomics. However, so far, no large-scale analysis of gene expression exists for this species. In the present study, gene expression was analyzed in gilthead sea bream during early development, a significant period in the determination of quantitative traits and therefore of considerable interest for aquaculture. Synexpression groups expressed primarily early and late in development were determined and were composed of both known and novel genes. Furthermore, it was possible to identify stress response genes induced by cortisol injections using the cDNA microarray generated. The creation of gene expression profiles for sea bream by microarray hybridization will accelerate identification of candidate genes involved in multifactorial traits and certain regulatory pathways and will also contribute to a better understanding of the genetic background of fish physiology, which may help to improve aquaculture practices.
- A gene-based radiation hybrid map of the gilthead sea bream Sparus aurata refines and exploits conserved synteny with Tetraodon nigroviridisPublication . Sarropoulou, Elena; Franch, Rafaella; Louro, Bruno; Power, Deborah; Bargelloni, Luca; Magoulas, Antonio; Senger, Fabrice; Kotoulas, Georgios; Geisler, RobertBackground: Comparative teleost studies are of great interest since they are important in aquaculture and in evolutionary issues. Comparing genomes of fully sequenced model fish species with those of farmed fish species through comparative mapping offers shortcuts for quantitative trait loci (QTL) detections and for studying genome evolution through the identification of regions of conserved synteny in teleosts. Here a comparative mapping study is presented by radiation hybrid (RH) mapping genes of the gilthead sea bream Sparus aurata, a non-model teleost fish of commercial and evolutionary interest, as it represents the worldwide distributed species-rich family of Sparidae. Results: An additional 74 microsatellite markers and 428 gene-based markers appropriate for comparative mapping studies were mapped on the existing RH map of Sparus aurata. The anchoring of the RH map to the genetic linkage map resulted in 24 groups matching the karyotype of Sparus aurata. Homologous sequences to Tetraodon were identified for 301 of the gene-based markers positioned on the RH map of Sparus aurata. Comparison between Sparus aurata RH groups and Tetraodon chromosomes (karyotype of Tetraodon consists of 21 chromosomes) in this study reveals an unambiguous one-to-one relationship suggesting that three Tetraodon chromosomes correspond to six Sparus aurata radiation hybrid groups. The exploitation of this conserved synteny relationship is furthermore demonstrated by in silico mapping of gilthead sea bream expressed sequence tags (EST) that give a significant similarity hit to Tetraodon. Conclusion: The addition of primarily gene-based markers increased substantially the density of the existing RH map and facilitated comparative analysis. The anchoring of this gene-based radiation hybrid map to the genome maps of model species broadened the pool of candidate genes that mainly control growth, disease resistance, sex determination and reversal, reproduction as well as environmental tolerance in this species, all traits of great importance for QTL mapping and marker assisted selection. Furthermore this comparative mapping approach will facilitate to give insights into chromosome evolution and into the genetic make up of the gilthead sea bream.
- The first radiation hybrid map of a perch-like fish: the gilthead seabream (Sparus aurata L)Publication . Senger, Fabrice; Priat, Catherine; Hitte, Christophe; Sarropoulou, Elena; Franch, Rafaella; Geisler, Robert; Bargelloni, Luca; Power, Deborah; Galibert, F.Among Teleosts, Perciformes are the largest order of fishes and include numerous species of commercial importance. Perciformes also comprise species of primary interest for evolutionary studies and analysis of the sex determination systems and sex chromosome plasticity. Unfortunately, genomics tools and resources for Perciformes remain to be developed. Here, we report the production of a seabream whole-genome radiation hybrid (RH) panel in which quality was ascertained by the construction of a 2-Mb-resolution RH map. The map encompasses 440 markers (288 microsatellites, 82 gene-based markers, and 70 STS) suitable for linkage analysis and comparative mapping studies. Achievement of a RHpanel and a whole-genome RH map should contribute to establishing seabream as a fish model among the Perciformes and should be of importance in aquaculture for marker-assisted selection, improvement of growth performance, and disease management. Development of RH maps in a costeffective manner for other fishes with the described methodology will offer a powerful approach in aquaculture and will provide extended capabilities for comparing vertebrate genome evolution.