FunnyBase: a systems level functional annotation of Fundulus ESTs for the analysis of gene expression.
BACKGROUND
While studies of non-model organisms are critical for many research areas, such as evolution, development, and environmental biology, they present particular challenges for both experimental and computational genomic level research. Resources such as mass-produced microarrays and the computational tools linking these data to functional annotation at the system and pathway level are rarely available for non-model species. This type of "systems-level" analysis is critical to the understanding of patterns of gene expression that underlie biological processes.


RESULTS
We describe a bioinformatics pipeline known as FunnyBase that has been used to store, annotate, and analyze 40,363 expressed sequence tags (ESTs) from the heart and liver of the fish, Fundulus heteroclitus. Primary annotations based on sequence similarity are linked to networks of systematic annotation in Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) and can be queried and computationally utilized in downstream analyses. Steps are taken to ensure that the annotation is self-consistent and that the structure of GO is used to identify higher level functions that may not be annotated directly. An integrated framework for cDNA library production, sequencing, quality control, expression data generation, and systems-level analysis is presented and utilized. In a case study, a set of genes, that had statistically significant regression between gene expression levels and environmental temperature along the Atlantic Coast, shows a statistically significant (P < 0.001) enrichment in genes associated with amine metabolism.


CONCLUSIONS
The methods described have application for functional genomics studies, particularly among non-model organisms. The web interface for FunnyBase can be accessed at http://genomics.rsmas.miami.edu/funnybase/super_craw4/. Data and source code are available by request at jpaschall@bioinfobase.umkc.edu.
Paschall
Justin E
JE
Division of Molecular Biology and Biochemistry, 5100 Rockhill Rd., University of Missouri, Kansas City 64110, USA.
Oleksiak
Marjorie F
MF
VanWye
Jeffrey D
JD
Roach
Jennifer L
JL
Whitehead
J Andrew
JA
Wyckoff
Gerald J
GJ
Kolell
Kevin J
KJ
Crawford
Douglas L
DL
D000818:Animals; D019295:Computational Biology / Q000379:methods; D018076:DNA, Complementary / Q000378:metabolism; D030541:Databases, Genetic; D020224:Expressed Sequence Tags; D005399:Fishes; D023781:Fundulidae / Q000235:genetics*; D015870:Gene Expression; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015723:Gene Library; D016678:Genome; D023281:Genomics; D020407:Internet; D005810:Multigene Family; D020411:Oligonucleotide Array Sequence Analysis; D012984:Software; D013696:Temperature
D018076:DNA, Complementary