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| Cloning and Derived Antibacterial Peptides of Histone H2A Gene in Black Carp Mylopharyngodon piceus,Grass Carp Ctenopharyngodon idellus,Silver Carp Hy pophthalmichthys molitrix and Bighead Carp Aristichthys nobilis |
| WU Xiang-min,ZHAO Yan-jing,KONG Xiang-hui,JIANG Hong-xia,LI Li,NIE Guo-xing,LI Xue-jun |
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Abstract Antimicrobial peptide derived from Histone H2A ,one of the chromosome structure protein ,is a cationic antimicrobial peptide showing a potential to substitute antibiotics .The characterization of histone H2A and the derivative antibacterial peptides are important for understanding of the mechanism and antibacterial difference among peptides derived from different H 2A genes in different fish .In this study , RT‐PCR was used to amply the H2A cDNA with the total RNA as the template in black carp Mylopharyngodon piceus , grass carp Ctenopharyngodon idellus , silver carp Hypophthalmichthys molitrix and bighead carp A ristichthys nobilis .The homologous H2A gene sequences were selected in the BLAST in GenBank ,and aligned in software Mega5 . The universal primers were designed in software Primer5 to amplify H2A genes collected and linked with a T‐vector ,and transferred into the competent cells for cloning .The H2A gene sequences with the length of 364 bp in the four fish species were cloned , and the deduced proteins were found to be unstable ,with the PI value of 10 .48—11 .02 and the total average hydrophilicity of 0 .179—0 .264 by online analysis . Both sequence homology analysis and the constructed phylogenetic tree revealed that the deduced H2A proteins showed the high homology (over 85% ) in four fish species with those in Sinocyclocheilus angustiporus and zebrafish .Histone H2A‐derived antibacterial peptide located in N terminal region in the four fish species showed the high homology with Hipposin in Atlantic halibut H ip poglossus hip poglossus .In addition ,there were glutamic acid residues in all amino acid sequences . It was confirmed that the antibacterial peptides derived from H 2A include Hipposin ,Parasin Ⅰ ,and Buforin ,belonging to cationic peptides with αspiral structure .
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Received: 25 May 2014
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