Gene Cloning and Expression of Crustin 6 in Procambarus clarkii

  Objective  Functions of crustacean gene of Procambarus clarkia were studied.

  Method  Pc-crustin6 was targeted for this study with specifically designed primers for PCR. Amino acids alignment and phylogenetic tree analysis were performed with bioinformatics software to decipher the molecular properties of Pc-crustin 6. Expressions and distributions of Pc-crustin 6 in different tissues were analyzed by qRT-PCR. The expression after an Edwardsiella ictaluri challenge was used to determine the disease resistance associated with the gene.

  Result  The full length Pc-crustin 6 cDNA was 465 bp with an open reading frame of 384 bp and encoding 127 amino acids. The N terminal of the molecule contained a signal peptide of 25 amino acids, and the C terminal a WAP domain of 8 conserved cysteine residues. Pc-crustin 6 was expressed to varied extents in 5 organs including the gills, which was the highest among them. After an artificial exposure to E. ictaluri, the expressions of Pc-crustin 6 in the hemocytes, hepatopancreas, gills, and intestines of P. clarkia were significantly up-regulated.

  Conclusion  The Pc-crustin 6 expression in organs of P. clarkia was significantly altered after being stimulated by the induction of E. ictaluri. It indicated that the gene was involved in the pathogenic resistance of P. clarkia.