Molecular cloning and sequencing of hemoglobin-beta gene of channel catfish, Ictalurus punctatus Rafinesque

Abstract

The hemoglobin-beta gene of channel catfish, Ictalurus punctatus, was cloned and sequenced. Total RNA from head kidneys was isolated, reverse transcribed and amplified. The sequence of the channel catfish hemoglobin-beta gene consists of 600 nucleotides. Analysis of the nucleotide sequence reveals one open reading frame and 5'- as well as 3'-untranslated regions. The open reading frame of the sequence potentially encodes 148 amino acids with a calculated molecular mass of 16.3 kDa. The pI and charge at pH 7.0 of the deduced hemoglobin-beta protein were 7.28 and 0.47, respectively. Overall, 22 amino acid residues were conserved throughout the sequences, including His64 and His93, the sites for heme-binding. Unlike the counterpart of other common cultured fish such as Salmo salar, Oncorhynchus nerka, Oncorhynchus mykiss, Cyprinus carpio and Ctenopharyngodon idella, the hemoglobin-beta of channel catfish did not have cysteine. The amino acid sequence of channel catfish hemoglobin-beta shows 84% homology with that of Silurus asotus (both are in the order Siluriformes). However, comparison with those of other fish species shows homology ranging from 53 to 68%. Structural analysis by the 3D-PSSM program displays that channel catfish hemoglobin-beta has eight alpha-helices, A-H.