To
date, no good breeding varieties have been obtained using the oriented cultivation technique.
The ability to identify disease resistance genes in grass carp is important to cultivate diseaseresistant varieties of grass carp.
In this study, we constructed a non-normalized cDNA library of head kidney in grass carp,
and, after clustering and assembly, we obtained 3,027 high-quality unigenes. Solexa
sequencing was used to generate sequence tags from the transcriptomes of the head kidney in
grass carp before and after grass carp reovirus (GCRV) infection.
After processing, we
obtained 22,144 tags that were differentially expressed by more than 2-fold between the
uninfected and infected groups. 679 of the differentially expressed tags (3.1%) mapped to 483 of the unigenes (16.0%).
The up-regulated and down-regulated unigenes were annotated using gene ontology terms; 16 were annotated as immune-related and 42 were of unknown
function having no matches to any of the sequences in the databases that were used in the
similarity searches.
Semi-quantitative RT-PCR revealed four unknown unigenes that showed
significant responses to the viral infection. Based on domain structure predictions, one of
these sequences was found to encode a protein that contained two transmembrane domains
and, therefore, may be a transmembrane protein. Here, we proposed that this novel unigene
may encode a virus receptor or a protein that mediates the immune signalling pathway at the
cell surface.
Conclusion
This study enriches the molecular basis data of grass carp and further confirms that, based on fish tissue-specific EST databases, transcriptome analysis is an effective route to discover novel functional genes.
October 2012 |
Further ReadingYou can view the full report and list of authors by clicking here. |
