Molecular cloning, characterization and expression analysis of complement component C8 beta in the orange-spotted...

Complement component C8 beta was a key molecule in the complement system, mediating the MAC formation and the bacterial lysis. In this study, the full-length C8 beta (EcC8 beta) was obtained, containing a 5'UTR of 25 bp, an ORF of 1764 bp and a 3'UTR of 198 bp. The EcC8 beta gene encoded a protein of 587 amino acids with an estimated molecular mass of 65.87 kDa and a predicted isoelectric point (pI) of 6.35. The deduced amino acid sequence showed that EcC8 beta consisted of the conserved residues and the domains known to be critical for C8 beta function. The quantitative real-time PCR analysis revealed that EcC8 beta transcript was expressed in all the examined tissues, while the strong expression was observed in the liver. In addition, complement C3 was the central molecule in the complement system, converging the upstream complement signals and mediating the MAC assembly pathway, while C8 beta was indispensable for active MAC formation. Following the Vibrio challenge, the increased expression of EcC3 transcript and EcC8 beta transcript was observed in the liver and kidney. These results indicated that EcC8 beta may be an important immune-related gene, playing an important role in the immune defense against the bacterial infection via the complement pathway.