DOI: 10.3724/SP.J.1118.2013.01188

Journal of Fishery Sciences of China (中国水产科学) 2013/20:6 PP.1188-1196

Gene characterization and expression of defensin in Cristaria plicata

The specimen of Cristaria plicata with shell length(9.50±1.05) cm and shell height(6.45±0.81) cm were temporally cultured for one week. We designed test trial and control trial with 20 individuals for each. We obtained the full-length cDNA sequence of defensin from the cDNA library of Cristaria plicata. The full-length cDNA was 514 bp, consisting of a 5'-terminal untranslated region(UTR)of 57 bp, a 3'-terminal UTR of 242 bp with a poly(A)tail, and an open reading frame of 192 bp which encoded a polypeptide of 63 amino acids residues, including 23 signal peptides and 40 mature peptides. The predicted molecular weight was 7.13 kD, the theoretical isoelectric point was 8.72. The predicted structure revealed that defensin gene contained a conserved cysteine-stabilized α-helix and two β-sheet(CSαβ)structural motif. We measured the expression defensin in the hemocytes, mantle, gill, and hepatopancreas of C. plicata by semi-quantitative RT-PCR following stimulation by Aeromonas hydrophila(109 cell/mL). Before stimulation, the expression of defensin was highest in the mantle and was about 1.25 times that in hemocytes and gill. After injection(at 0, 6, 12, 24, 36, 48 h), the relative expression first decreased in the mantle and hepatopancreas then increased significantly in the control group. There was no change in defensin expression in the hemocytes and gill. In the experimental group, expression peaked after 12 h in the hemocytes then decreased. Conversely, defensin expression was highest in the other tissues at 0 h, and then decreased gradually. Expression increased slightly in the mantle and gill at 24 h and 36 h.

Key words:Cristaria plicata,defensin,gene expression,sequence structure,mytilin,Aeromonas hydrophila

ReleaseDate:2014-07-21 16:58:30