Molecular cloning and expression analysis of a CXC chemokine gene from large yellow croaker Pseudosciaena crocea

In the present study, we report the cloning of a CXCL12 chemokine gene homologue from the large yellow croaker Pseudosciaena crocea (LycCXCL12). The complete cDNA of LycCXCL12 is 678 nucleotides (nt) encoding a protein of 97 amino acids (aa), with a putative molecular weight of 11.1 kDa. The deduced LycCXCL12 contains a 22-aa signal peptide and a 75-aa mature polypeptide, which possesses the typical arrangement of four cysteines as found in other known CXC chemokines. It shares 57–68% and 32–36% aa sequence identities to known CXCL12 chemokines in fish species and other vertebrates, respectively. The LycCXCL12 gene was constitutively expressed in all tissues examined although at different levels. Upon induction with poly(I:C) or inactivated trivalent bacterial vaccine, LycCXCL12 gene expression was significantly up-regulated in gills, liver, kidney, spleen and blood at 24 h after stimulation. Time course analysis using real-time PCR showed that LycCXCL12 gene expression reached peak level in spleen and kidney at 12 h or in gills at 24 h post-induction by poly(I:C), while its expression increased to the highest level in kidney at 24 h or in gills and spleen at 48 h post-induction by bacterial vaccine, indicating that LycCXCL12 gene expression was differentially regulated by poly(I:C) and bacterial vaccine.