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Gene cloning, expression and characterization of avian cathelicidin orthologs, Cc‐CATHs, from Coturnix coturnix
Author(s) -
Feng Feifei,
Chen Chen,
Zhu Wenjuan,
He Weiyu,
Guang Huijuan,
Li Zheng,
Wang Duo,
Liu Jingze,
Chen Ming,
Wang Yipeng,
Yu Haining
Publication year - 2011
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2011.08080.x
Subject(s) - cathelicidin , biology , antimicrobial , orfs , cloning (programming) , microbiology and biotechnology , gene , antimicrobial peptides , recombinant dna , open reading frame , genetics , peptide sequence , computer science , programming language
Cathelicidins comprise a family of antimicrobial peptides sharing a highly conserved cathelin domain, which play a central role in the early innate host defense against infection. In the present study, we report three novel avian cathelicidin orthologs cloned from a constructed spleen cDNA library of Coturnix coturnix , using a nested‐PCR‐based cloning strategy. Three coding sequences containing ORFs of 447, 465 and 456 bp encode three mature antimicrobial peptides (named Cc‐CATH1, 2 and 3) of 26, 32 and 29 amino acid residues, respectively. Phylogenetic analysis indicated that precursors of Cc‐CATHs are significantly conserved with known avian cathelicidins. Synthetic Cc‐CATH2 and 3 displayed broad and potent antimicrobial activity against most of the 41 strains of bacteria and fungi tested, especially the clinically isolated drug‐resistant strains, with minimum inhibitory concentration values in the range 0.3–2.5 μ m for most strains with or without the presence of 100 m m NaCl. Cc‐CATH2 and 3 showed considerable reduction of cytotoxic activity compared to other avian cathelicidins, with average IC 50 values of 20.18 and 17.16 μ m , respectively. They also exerted a negligible hemolytic activity against human erythrocytes, lysing only 3.6% of erythrocytes at a dose up to 100 μg·mL −1 . As expected, the recombinant Cc‐CATH2 (rCc‐CATH2) also showed potent bactericidal activity. All these features of Cc‐CATHs encourage further studies aiming to estimate their therapeutic potential as drug leads, as well as coping with current widespread antibiotic resistance, especially the new prevalent and dangerous ‘superbug’ that is resistant to almost all antibiotics.