Expressed sequence tags from normalized cDNA libraries prepared from gill and hypodermal tissues of the blue crab,...

Expressed sequence tags (ESTs) were produced from two normalized cDNA libraries from the blue crab, Callinectes sapidus. The gill library represented pooled RNA from respiratory and transporting gills after acclimation to either high or low salinity. The hypodermis library was from arthrodial and dorsal tissue from both pre- and post-molt crabs. Random clones were single-pass sequenced from the 5'-ends, resulting in 11,761 high quality ESTs averaging 652 bases. All the ESTs were assembled using Paracel Transcript Assembler software, producing 2176 potential transcripts-883 contigs and 1293 singlets. Of these, 1235 (56.7%) were sequenced only from the gill library, while 578 (26.6%) were exclusively hypodermal. There were 363 contigs containing ESTs from both tissues (16.7% of the putative transcripts). All contigs and singlets were compared to the public protein database using BLASTx, and descriptions of the three most similar proteins for each were recorded. Additional annotations included an Interpro analysis of protein domains and a listing of Gene Ontology (GO) categories inferred from similar proteins in GO-annotated databases. All sequences are available on a web page (http://firedev.bear.uncw.edu:8080/shaferlab/). The annotations can be searched, and BLAST alignment of user-inputted sequences against the putative transcripts is possible. In addition, the ESTs have been submitted to GenBank.