Full-length cDNA sequence of 14-3-3 gene was isolated from the eyestalk of mud crab Scylla paramamosain through RT-PCR and RACE. Sequence analysis indicated that 14-3-3 gene had an open reading frame of 744 bp encoding 247aa of 28.086 ku and pI at 4.675. The amino acid sequences of 14-3-3 gene possessed 95%, 93%, 92% identity with the 14-3-3 genes of Penaeus monodon, Fenneropenaeus merguiensis, Megachile rotundata respectively. 14-3-3 protein firstly clustered with 14-3-3 proteins of Penaeus monodon and Fenneropenaeus merguiensis in the phylogenetic analysis. The expression of 14-3-3 gene in tissues was analyzed by Real-Time PCR, the result showed that 14-3-3 gene was most expressed in hepatopancreas and muscle, then in eyestalk, intestinal, heart and gill, the lest in stomach. After 24h’s stress in salinity, the expression of 14-3-3 gene increased great significantly (P<0.01) whatever the salinity reduced (5) or raise (15, 20, 25, 30). The more the salinity changed, the more the 14-3-3 gene expressed. These results will serve further studies on functions and regulation mechanism of 14-3-3 genes.