title: Identification, structural characterisation and expression analysis of a defensin gene from the tiger beetle Calomera littoralis (Coleoptera: Cicindelidae).
abstract: In this study, a defensin gene (Clit-Def) has been characterised in the tiger beetle Calomera littoralis for the first time. Bioinformatic analysis showed that the gene has an open reading frame of 246bp that contains a 46 amino acid mature peptide. The phylogenetic analysis showed a high variability in the coleopteran defensins analysed. The Clit-Def mature peptide has the features to be involved in the antimicrobial function: a predicted cationic isoelectric point of 8.94, six cysteine residues that form three disulfide bonds, and the typical cysteine-stabilized α-helix β-sheet (CSαβ) structural fold. Real time quantitative PCR analysis showed that Clit-Def was upregulated in the different body parts analysed after infection with lipopolysaccharides of Escherichia coli, and also indicated that has an expression peak at 12h post infection. The expression patterns of Clit-Def suggest that this gene plays important roles in the humoral system in the adephagan beetle Calomera littoralis.
authors.affiliation: University of Murcia, Department of Zoology and Physical Anthropology, Faculty of Veterinary, Campus Mare Nostrum, E-30100 Murcia, Spain. Electronic address: mjulia.rodriguez@um.es.
authors.forename: María Juliana
authors.initials: MJ
authors.lastname: Rodríguez-García
authors.affiliation: University of Murcia, Department of Zoology and Physical Anthropology, Faculty of Veterinary, Campus Mare Nostrum, E-30100 Murcia, Spain.
authors.forename: Andrés
authors.initials: A
authors.lastname: García-Reina
authors.affiliation: University of Murcia, Department of Zoology and Physical Anthropology, Faculty of Veterinary, Campus Mare Nostrum, E-30100 Murcia, Spain.
authors.forename: Vilmar
authors.initials: V
authors.lastname: Machado
authors.affiliation: University of Murcia, Department of Zoology and Physical Anthropology, Faculty of Veterinary, Campus Mare Nostrum, E-30100 Murcia, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Galián
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001517:Coleoptera / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D019295:Computational Biology; D023082:Defensins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry / Q000502:physiology; D005786:Gene Expression Regulation; D007526:Isoelectric Point; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology; D016366:Open Reading Frames; D010802:Phylogeny; D017433:Protein Structure, Secondary; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D023082:Defensins; D004220:Disulfides; D008070:Lipopolysaccharides
keywords: AMP; Adephagan beetles; Cicindelids; Defensin; RT-qPCR