Cloning and expression analysis of 14-3-3 gene in Portunus trituberculatus after exposure to low salt and pathogenic stress

Abstract:

In this study, the 14-3-3 gene of the swimming crab (Portunus trituberculatus) was cloned using the Rapid Amplification of cDNA Ends (RACE) method. The full length was 2510 bp and the ORF was 741 bp, encoding 247 amino acids and with a predicted molecular weight of 27.98 kDa. Sequence alignment analysis showed that the Pt14-3-3 gene has the highest homology with the 14-3-3 gene of Eriocheir sinensis (100%). Phylogenetic tree analysis indicated that the Pt14-3-3 amino acid sequence of P. trituberculatus was closely clustered into one with Eriocheir sinensis. Tissue expression analysis showed that the Pt14-3-3 gene was expressed in hepatopancreas, muscle, gill, heart, eyestalk, and hemolymph, and the expression level was highest in hepatopancreas. After low salt stress, the expression levels of the Pt14-3-3 gene in sputum and hepatopancreas were significantly upregulated at 48 h and 12 h, respectively, and reached the maximum, which was 1.34 times (P<0.05) and 7.54 times (P<0.05), respectively. After artificial infection with Vibrio parahaemolyticus and WSSV, the Pt14-3-3 gene was upregulated in the hepatopancreas and blood cells and the lowest was up to 17.52 times (P<0.05). The results of this study indicated that the Pt14-3-3 gene plays an important role in low salt adaptation and immune response of the Portunus trituberculatus.