研究简报 Short Communications

棘腹蛙Japonicin-pb抗菌肽的分离及表达谱分析

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  • 1. 重庆文理学院林学与生命科学学院, 重庆 402160;
    2. 重庆珍稀濒危水产资源保护与开发研究中心, 重庆 402168
姜玉松(1984—),男,山东高密人,讲师,博士,从事基因的分子生物学研究。E-mail:jysong@126.com

收稿日期: 2014-12-23

  网络出版日期: 2015-05-13

基金资助

重庆市科技攻关计划(cstc2012gg-yyjs80004);重庆市教委科学技术研究项目(KJ1401105);重庆永川区自然基金项目(Ycstc,2014nc1001);重庆文理学院人才引进项目(R2014LX07,R2013LS13)

Isolation and Expression Patterns of Japonicin-pb from Paa boulengeri

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  • 1. College of Life Science & Forestry, Chongqing University of Art & Science, Chongqing 402160, China;
    2. Chongqing Research Centers of Conservation & Development on Rare & Endangered Aquatic Resources, Chongqing 402168, China

Received date: 2014-12-23

  Online published: 2015-05-13

摘要

本试验旨在克隆棘腹蛙来源Japonicin-pb,了解该抗菌肽在不同生长温度及组织下的表达规律,为Japonicin-pb的开发提供线索。基于本实验室构建的棘腹蛙皮肤转录组数据库,利用反转录PCR从棘腹蛙皮肤组织中克隆Japonicin-pb前体序列,通过结构域比对的方法分析其结构。利用Real-time PCR检测3种Japonicin-pb在不同生长温度(15、18、21、24、27和30 ℃)及组织(血液、肌肉、肝脏和皮肤)的表达图谱。结果表明,本试验克隆到3种Japonicin-pb前体序列,结构域比对表明,三者拥有相同的N端信号肽和中间间隔区,长度分别为19、20和12个氨基酸残基的肽序列,命名为Japonicin-1apb、Japonicin-1bpb和Japonicin-3pb。Japonicin-1apb主要在皮肤组织中表达,且表达量随着生长温度的升高而逐渐增加;Japonicin-1bpb在不同组织或生长温度下的表达量差异均不明显;Japonicin-3pb主要在肝脏中表达,且生长温度对其影响不大。这些结果为棘腹蛙源3种Japonicin-pb后续功能研究以及开发应用提供了重要的序列及表达谱信息。

本文引用格式

姜玉松, 陈德碧, 邹勇, 周继萍, 樊汶樵, 徐敬明 . 棘腹蛙Japonicin-pb抗菌肽的分离及表达谱分析[J]. 动物营养学报, 2015 , 27(5) : 1613 -1619 . DOI: 10.3969/j.issn.1006-267x.2015.05.034

Abstract

In order to provided clues to the development of Japonicin-pb, we cloned the Japonicin-pb from Paa boulengeri and investigated its expression patterns among different growth temperatures and tissues. The complete open reading frame (ORF) sequence of Japonicin-pb precursor was amplified by RT-PCR according to the transcriptome database of Paa boulengeri constructed by our laboratory. The sequential structure of Japonicin-pb was analyzed by the method of domain alignment. The expression patterns of Japonicin-pb among different growth temperatures (15, 18, 21, 24, 27 and 30 ℃) and tissues (blood, muscle, liver and skin) were analyzed by real-time PCR. The results showed that according to the domain alignment, 3 Japonicin-pb precursors had the same N terminal signal peptide, middle spacer region, but the mature peptides were 19, 20 and 12 amino acid residues, named Japonicin-1apb, Japonicin-1bpb and Japonicin-3pb, respectively. The expression patterns showed that Japonicin-1apb was mainly expressed in skin, and its expression abundance increased gradually from 15 to 30 ℃; the expression level of Japonicin-1bpb among different tissues and growth temperatures were no significant difference; Japonicin-pb mainly distributed in liver, and it was not sensitive to the response of growth temperature. These results provide important clues for the functional study and the development of Japonicin-pb.

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