CLONING AND EXPRESSION OF WHITE CLOUD MOUNTAIN MINNOW(TANICHTHYS ALBONUBES) CASPASE-3 AND CASPASE-9 cDNAs

The white cloud mountain minnow(Tanichthys albonubes) is an important aquarium fish,and also a potential experimental model organism.In order to get the knowledge of apoptosis under environmental stress in this species,we have firstly cloned and sequenced the full-length cDNAs of Caspase-3 and Caspase-9 from the gill tissue of T.albo-nubes using homology cloning approach combined with RACE method.In this study,the bioinformatics analysis of Caspase-3 and Capase-9 full-length cDNA sequences were presented.The results indicated that the full-length cDNA of Caspase-3 was composed of 2153 base pairs(bp),which included a 840 bp open reading frame(ORF) encoding a 279-amino acid peptide,whereas the full-length cDNA of Caspase-9 was composed of 1758 bp,which included a 1323 bp ORF encoding a 440-amino acid peptide.The identity comparison and the phylogenetic analysis showed that the predicted Caspase-3 and Caspase-9 proteins of T.albonubes were structurally similar to those of other fish,with high conservation in important functional domains.Real-time PCR analysis showed that the expression levels of Caspase-3 and Caspase-9,under cypermethrin-induced stress conditions,were suppressed compared to control group,and that Caspase-3 and Caspase-9 mRNA levels were significantly affected in a dose-and time-dependent manner by cyperme-thrin.These results suggest that cypermethrin plays a role in the suppression of apoptosis of T.albonubes,and that the anti-apoptotic effect of cypermethrin may be beneficial to this fish so that it can survive in harsh environments.