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首页> 外文期刊>Fish & Shellfish Immunology >Molecular, genomic, and expressional delineation of a piscidin from rock bream (Oplegnathus fasciatus) with evidence for the potent antimicrobial activities of Of-Pis1 peptide
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Molecular, genomic, and expressional delineation of a piscidin from rock bream (Oplegnathus fasciatus) with evidence for the potent antimicrobial activities of Of-Pis1 peptide

机译：岩鲷（Oplegnathus fasciatus）中的甜菜碱的分子，基因组和表达描述，以及Of-Pis1肽的强效抗菌活性证据

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The piscidin family comprises a group of antimicrobial peptides (AMPs) that are vital components of teleost innate immunity. Piscidins protect the host from pathogens, through multifaceted roles as immunomodulators and anti-infective peptides. The present study reports the identification, and characterization of a putative piscidin homolog, Of-Pis1, from rock bream (Oplegnathus fasciatus). A combined genomic and transcriptomic approach revealed that the Of-Pis1 gene comprises 1396 nucleotides (nt), four exons, and three introns. The cDNA with the 213 nt open reading frame encoded a 70-amino acid preprotein consisting of a signal peptide, a mature peptide, and a prodomain. Predicted mature Of-Pis1 was assumed to be a membrane-active AMP, based on the prediction of an amphipathic alpha-helical conformation with a net charge of +4. In addition, Of-Pis1 demonstrated significant similarities with other piscidin family members in terms of gene structure, sequence homology, and evolutionary relationship. Examination by quantitative real-time PCR (qPCR) of basal transcription of Of-Pis1 in the tissues of naive rock bream, revealed predominant transcript levels in the gills, followed by the spleen, intestine, skin, and head kidney. In gill tissues, the temporally induced mRNA expression of Of-Pis1, upon in vivo injection trials with lipopolysaccharide (LPS); polyinosinic:polycytidylic acid (poly I:C); and pathogens, including Edwardsiella tarda, Streptococcus iniae, and rock bream iridovirus (RBIV), was weak. In contrast, in vivo flagellin administration led to a robust upregulation of Of-Pis1 in different tissues. Antimicrobial potency was determined by employing recombinant (rOf-Pis1), and synthetic (pOf-Pis1) peptides, in in vitro assays. Recombinant overexpression inhibited the growth of bacteria expressing the rOf-Pis1 protein in a growth delay assay. The broad antimicrobial spectrum of pOf-Pis1 was evidenced by its potent activity against an array of microbes, including bacteria, fungi, and parasitic species. In addition, pOf-Pis1 showed no significant hemolytic toxicity against human erythrocytes. Collectively, the data presented in the current study improve our understanding of the piscidin AMP family, and the contribution of Of-Pis1 to the rock bream immunity. (C) 2015 Elsevier Ltd. All rights reserved.
机译：piscidin系列包含一组抗菌肽（AMPs），它们是硬骨鱼先天免疫的重要组成部分。杀虫蛋白通过作为免疫调节剂和抗感染肽的多方面作用，保护宿主免受病原体侵害。本研究报告鉴定和表征的岩鲷（Oplegnathus fasciatus）的拟南芥同源物Of-Pis1。基因组和转录组学研究相结合的方法显示，Of-Pis1基因包含1396个核苷酸（nt），四个外显子和三个内含子。具有213nt开放阅读框的cDNA编码由信号肽，成熟肽和前结构域组成的70个氨基酸的前蛋白。基于对净电荷为+4的两亲性α螺旋构象的预测，假定预测的Of-Pis1成熟为膜活性AMP。此外，Of-Pis1在基因结构，序列同源性和进化关系方面与其他piscidin家族成员表现出显着相似性。通过定量实时PCR（qPCR）对幼稚rock鱼组织中Of-Pis1的基础转录的检查，揭示了g中主要的转录水平，其次是脾脏，肠，皮肤和头肾。在g组织中，脂多糖（LPS）体内注射试验中，时间诱导的Of-Pis1 mRNA表达;聚肌苷酸：聚胞苷酸（poly I：C）;病原体包括爱德华氏菌，印度链球菌和鲷鱼虹膜病毒（RBIV）都很弱​​。相反，体内鞭毛蛋白施用导致不同组织中的Of-Pis1强烈上调。在体外测定中，通过使用重组（rOf-Pis1）和合成（pOf-Pis1）肽来确定抗菌效力。在生长延迟试验中，重组过表达抑制了表达rOf-Pis1蛋白的细菌的生长。 pOf-Pis1对多种微生物（包括细菌，真菌和寄生物）具有有效的活性，证明了其广泛的抗菌谱。此外，pOf-Pis1对人红细胞没有明显的溶血毒性。总的来说，当前研究中提供的数据增进了我们对piscidin AMP系列的了解，以及Of-Pis1对鱼免疫力的贡献。 （C）2015 Elsevier Ltd.保留所有权利。

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