棘腹蛙皮肤抗菌肽Cathelicidin-Pb的克隆及表达分析  

Cloning and expression analysis of Cathelicidin-Pb from Paa boulengeri

作　　者：姜玉松[1,2] 张文琪[1] 樊汶樵[1,2] 徐敬明[1,2] 

机构地区：[1]重庆文理学院林学与生命科学学院,重庆永川402160 [2]重庆珍稀濒危水产资源保护与开发研究中心,重庆永川402168

出　　处：《淡水渔业》2015年第4期26-30,共5页Freshwater Fisheries

基　　金：重庆文理学院人才引进项目(R2014LX07;R2013LS13);永川区自然科学基金项目(Ycstc;2014nc1001);重庆市科技攻关计划(cstc2012gg-yyjs80004)

摘　　要：基于棘腹蛙(Paa boulengeri)皮肤转录组数据库,釆用RT-PCR方法从棘腹蛙皮肤组织中克隆到了编码Cathelicidin-Pb前体的核苷酸序列,其完整ORF长度为441 bp,编码146个氨基酸残基。通过信号肽预测和结构域查找比对,发现Cathelicidin-Pb前体包含N端20个氨基酸残基的信号肽、101个氨基酸残基的中间间隔区(cathelin结构域)和25个氨基酸残基的成熟抗菌肽。进一步研究发现,在适度高温范围内,皮肤组织中Cathelicidin-Pb的表达随着生长环境温度的升高而逐渐增加,但是当环境温度超过27℃后,其表达量急剧降低;Cathelicidin-Pb主要在皮肤组织中表达,在肌肉组织和血液组织的表达量较低。这些结果为棘腹蛙养殖过程中疾病控制及Cathelicidin-Pb抗菌肽的后续开发提供了重要的线索。Based on the transcriptome database of Paa boulengeri, primers were designed to clone the complete open read-ing frame （ ORF） sequence of Cathelicidin-Pb precursor gene by RT-PCR.The result showed that the ORF of Cathelicidin-Pb precursor was 441 bp and encoded 146 amino acid residues.Cathelicidin-Pb precursor consisted of three parts, inclu-ding an N-terminal signal peptide sequence of 20 amino-acid residues, a middle spacer region of 101 amino acid residues （ Cathelin domain） and a mature peptide sequence of 25 amino acid residues.Further study found that, the expression of Cathelicidin-Pb in skin tissue, increased gradually along with the increase of environmental temperature, but when the en-vironment temperature above 27 ℃, the expression amount decreased dramatically.In addition, compared with muscle and blood tissue Cathelicidin-Pb is mainly expressed in skin tissue.These results provide important clues for the disease control in Paa boulengeri breeding process and for the development of antimicrobial peptides.

关 键 词：棘腹蛙(Paa boulengeri) 抗菌肽 差异表达 

分 类 号：S917.4[农业科学—水产科学]