CLONING AND EXPRESSION OF WHITE CLOUD MOUNTAIN MINNOW (TANICHTHYS ALBONUBES) CASPASE-3 AND CASPASE-9 cDNAs
唐鱼Caspase-3和Caspase-9 cDNA全长克隆及胁迫表达分析

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.