Abstract
BackgroundTo identify as many different transcripts/genes in the Atlantic salmon genome as possible, it is crucial to acquire good cDNA libraries from different tissues and developmental stages, their relevant sequences (ESTs or full length sequences) and attempt to predict function. Such libraries allow identification of a large number of different transcripts and can provide valuable information on genes expressed in a particular tissue at a specific developmental stage. This data is important in constructing a microarray chip, identifying SNPs in coding regions, and for future identification of genes in the whole genome sequence. An important factor that determines the usefulness of generated data for biologists is efficient data access. Public searchable databases play a crucial role in providing such service.DescriptionTwenty-three Atlantic salmon cDNA libraries were constructed from 15 tissues, yielding nearly 155,000 clones. From these libraries 58,109 ESTs were generated, of which 57,212 were used for contig assembly. Following deletion of mitochondrial sequences 55,118 EST sequences were submitted to GenBank. In all, 20,019 unique sequences, consisting of 6,424 contigs and 13,595 singlets, were generated. The Norwegian Salmon Genome Project Database has been constructed and annotation performed by the annotation transfer approach. Annotation was successful for 50.3% (10,075) of the sequences and 6,113 sequences (30.5%) were annotated with Gene Ontology terms for molecular function, biological process and cellular component.ConclusionWe describe the construction of cDNA libraries from juvenile/pre-smolt Atlantic salmon (Salmo salar), EST sequencing, clustering, and annotation by assigning putative function to the transcripts. These sequences represents 97% of all sequences submitted to GenBank from the pre-smoltification stage. The data has been grouped into datasets according to its source and type of annotation. Various data query options are offered including searches on function assignments and Gene Ontology terms. Data delivery options include summaries for the datasets and their annotations, detailed self-explanatory annotations, and access to the original BLAST results and Gene Ontology annotation trees. Potential presence of a relatively high number of immune-related genes in the dataset was shown by annotation searches.
Highlights
To identify as many different transcripts/genes in the Atlantic salmon genome as possible, it is crucial to acquire good cDNA libraries from different tissues and developmental stages, their relevant sequences (ESTs or full length sequences) and attempt to predict function
This paper describes a combined biological and bioinformatics resource and database dedicated to EST sequences from tissue specific pre-smolt Atlantic salmon cDNA libraries
Nineteen libraries were constructed from normal tissues and 4 libraries were constructed from gills and intestine using Suppression Subtractive Hybridisation (SSH) after intra-peritoneal injection of infectious salmon anaemia virus (ISAV) [15]
Summary
We have constructed 23 tissue specific cDNA libraries from pre-smolt Atlantic salmon (Salmo salar). 30.5% of sequences were further annotated using annotation transfer procedure for Gene Ontology (GO) terms for molecular function, biological process and cellular component. Annotation searches on biologically important putative functions showed that the [GO: immune response] term is over-represented in the SGP dataset, suggesting the presence of a relatively high number of immune-related genes. A database search, which can be done using sophisticated keywords search options, will produce an overview of the highly reliable sequence similarities ("best hits") and their gene and function annotations including GO assignments. AAA supervised SGP bioinformatics, designed software and data processing techniques and developed the data resource. He carried out software development and data processing and drafted part of the manuscript.
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