Molecular cloning and expression of 14-3-3ζ in Haliotis diversicolor under stresses

Tyrosine-3-monooxygenase/Tryp-tophane-5-monooxygenase activator protein(14-3-3ζ protein) is one of the isoform of 14-3-3 proteins which are highly conserved small acidic proteins.Several studies have shown that 14-3-3 proteins can bind many signal proteins, such as kinase and phosphatase, and play crucial roles in many biological processes including regulation of signaling and cell apoptosis.In this study, the full length cDNA of 14-3-3ζ from Haliotis diversicolor was cloned for the first time by SMART-RACE and named Hd14-3-3ζ.Its full-length cDNA sequence was 3 010 bp, with an open reading frame of 819 bp encoding a protein of 272 amino acids.The Real-time Quantitative PCR results showed that Hd14-3-3ζ was detected in all examined tissues and with the highest expression level in hepatopancreas and haemocytes.When under the stress of hypoxia, the expression level of Hd14-3-3ζ in haemocytes of exposed groups showed a significant up-regulation at 4 h, but in gill it happened at 24 h and 96 h.Under thermal stress, in haemocytes, it showed no significant difference between control and thermal groups but it was up-regulated significantly at 96 h in gill.Under thermal & hypoxia stresses, it showed no significant difference between control and exposed groups in haemocytes.However, in gill, it was up-regulated significantly at 4 h, 24 h and 96 h.After Vibrio parahaemolyticus challenge, the expression of Hd14-3-3ζ reached a significant level at 6 h in haemocytes and reduced to the normal level at 12 h, and then reached the highest significant level at 24 h.In gill, the significant expression level appeared at 3 h and 24 h and there was no difference between other time phases.All these results indicate that Hd14-3-3ζ may play important roles in immunoreaction of H.diversicolor.