The full length cDNA cloning and expression profile of 14-3-3 gene from the mud crab( Scylla paramamosain)

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(P0.01) 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.