Cloning and Prokaryotic Expression of Cathelicidin Gene from Japanese Eel I，Anguilla japonica

doi:10.13560/j.cnki.biotech.bull.1985.2015.07.018

Abstract

Abstract: Cathelicidin, an utmost family of antibacterial peptide so far, possesses multiple biological functions. In order to explore and exploit the antibacterial mechanism and potential biological functions of Cathelicidin, full-length sequence of cDNA of Cathelicidin gene from Japanese eel Anguilla japonica I(AjCathI)was obtained by RACE-PCR. The full cDNA of AjCathI was 842 bp and the ORF contained 570 bp encoding 189 amino acids. Analysis of amino acid sequence demonstrated that AjCathI contained 4 conservative cysteine residues at C terminal, and the mature antibacterial peptide had the highest similarity with Plecoglossus altivelis by homology of 65.57%. Phylogenetic analysis revealed that AjCathI shared a common evolutionary origin with other teleost fishes. AjCathI was cloned into pET-28a and expressed in Escherichia coli BL21(DE3). The result manifested that AjCathI protein was expressed in inclusion bodies. The protein was isolated by Ni column, identified by Western blot and re-natured by dialysis, the protein of high purity was gained. This study laid the foundation for further verifying mature peptide sequence of AjCathI and studying its antibacterial activities as well as other biological functions.

Key words: Anguilla japonica, Cathelicidin, antibacterial peptide, RACE-PCR

Zhang Dongling, Yu Dahui. Cloning and Prokaryotic Expression of Cathelicidin Gene from Japanese Eel I，Anguilla japonica[J]. Biotechnology Bulletin, 2015, 31(7): 124-131.

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