Molecular cloning and functional characterization of cyclophilin A in yellow catfish (Pelteobagrus fulvidraco)

Cyclophilin A (CypA) is a ubiquitously expressed protein which involves in diverse pathological conditions including infection and inflammation. In this report, a CypA gene (designated as YC-CypA) was cloned from yellow catfish (Pelteobagrus fulvidraco) which is an important cultured fish species in Asian countries. The open reading frame (ORF) of YC-CypA encoded a polypeptide of 164 amino acids with calculated molecular weight of 17.70 kDa. The deduced amino acid sequences of the YC-CypA shared highly conserved structures with CypAs from the other species, indicating that YC-CypA should be a new member of the CypA family. Full-length YC-CypA protein was expressed in Escherichia coli and specific polyclonal antibody against YC-CypA was generated. The YC-CypA protein showed chemotactic activity by transwell migration assay. The mRNA and protein of YC-CypA could be detected in all examined tissues with relatively higher mRNA level in spleen and higher protein level in head kidney, respectively. The temporal expression patterns of YC-CypA, IL-1 beta and TNF-alpha mRNAs were analyzed in the liver, spleen and head kidney post of Edwardsiella ictaluri infection. By immunohistochemistry assay, slight enhancement of YC-CypA protein was observed in the liver, spleen, body kidney and head kidney of yellow catfish infected with E. ictaluri. In conclusion, YC-CypA of yellow catfish showed chemotactic activity in vitro and might have been involved in cytokines secretion in yellow catfish during the infection of E. ictaluri. (C) 2015 Elsevier Ltd. All rights reserved.