[Cloning and expression analysis of Cathepsin L cDNA of Exopalaemon carinicauda]

Abstract

Based on the EST sequence from a hemocyte cDNA library, the cathepsin L cDNA of Exopalaemon carinicauda (EcCatL) was cloned by rapid amplification of cDNA ends (RACE). The EcCatL cDNA was 1136 bp in length, which contains an open reading frame (ORF) of 960 bp, encoding a 319 amino-acid polypeptide. Homology analysis revealed that the amino acid sequence of EcCatL was highly conserved with its homologs in other crustaceans. The similarities of EcCatL with the CatL of Palaemonetes varians and Pandalus borealis were 92% and 76%, respectively. Phylogenetic analysis showed that EcCatL was in the same branch as that of Palaemonetes varians. The expression levels of EcCatL in different tissues were analyzed by quantitative real-time PCR. Expression of EcCatL was detected in all tested tissues of E. carinicauda, including hemocytes, gill, hepatopancreas, muscle, ovary, intestine, stomach and eyestalk, with the highest expression level in hepatopancreas. After challenged with Vibrio anguillarum or white spot syndrome virus, the expression of EcCatL were up-regulated in the hemocytes and hepatopancreas of E. carinicauda. Our results implied that EcCatL might play an important role in the prawn immune response.

Keywords: Cathepsin L; Exopalaemon carinicauda; Gene cloning; Gene expression.