Molecular cloning and characterization of carp ( Cyprinus carpio L.) CD8β and CD4-like genes

Abstract

Partial cDNA sequences of both CD8β and CD4-like ( CD4L) genes of common carp ( Cyprinus carpio L.) were isolated from thymus cDNA library by the method of suppression subtractive hybridization (SSH). Subsequently the full length cDNAs of carp CD8β and CD4L were obtained by means of 3′ RACE and 5′ RACE, respectively. The full length cDNA of carp CD8β is 1164 bp and encodes 207 amino acids including a signal peptide region of 24 amino acids, a transmembrane region of 23 amino acids from aa 167 to aa189 and an immunoglobulin V-set from aa 19 to aa 141. Similar to other species CD8βs, carp CD8β also lacks p56 lck domain in the cytoplasmic region. The full length cDNA of carp CD4L is 2001 bp and encodes 458 amino acids including four immunoglobulin (Ig)-like domains in the extracellular region, a transmembrane region of 23 amino acids at the C-terminal region from aa 402 to aa 424 and a cytoplasmic tail. Similar to mammalian, avian CD4s and fugu CD4L, carp CD4L also has the conserved p56 lck tyrosine kinase motif (C-X-C) in the cytoplasmic region. RT-PCR analysis demonstrated that carp CD8β and CD4L genes were both expressed predominantly in thymus. The results from this study can be used to understand the evolution of both the CD8β and CD4 molecules which can be used as markers for cytotoxic and helper T cells in carp.