PDF《张毓霞大弹涂鱼皮肤转录组测序及抗菌肽基因分析_张毓霞》

大弹涂鱼皮肤转录组测序及抗菌肽基因分析 收稿日期: 2017-05-09;

修回日期: 2017-07-14 基金项目: 国家自然科学基金资助助项目(31472279) 作者简介: 张毓霞(1990-), 女, 山东临沂人, 硕士研究生, 主要从事海洋生物转录组和蛋白质组学研究;

* 通讯作者: 廖智(1973-), 男, 湖南 长沙人, 博士, 浙江海洋大学教授, 主要从事海洋生物蛋白质化学和蛋白质组学研究, E-mail: liaozhi@zjou.

edu.cn. 张毓霞1 , 石戈1 , 王日昕2 , 宫延斌1 , 范美华1 , 廖智1* (1. 浙江海洋大学 海洋科学与技术学院 海洋生物蛋白质工程实验室, 中国浙江 舟山 316022;

2. 宁波大学 海洋学院, 中国浙江 宁波 315211) 摘要: 抗菌肽是鱼类用于抵御外界微生物入侵的天然防御多肽, 也是开发新型药物的重要先导分子遥 大弹涂 鱼(Boleophthalmus pectinirostris)是一种特殊的可以营两栖生活的鱼类, 其皮肤表面具有丰富的粘液, 其中所含 抗菌肽对其免疫防御和适应两栖生活具有重要意义遥 为深入了解大弹涂鱼皮肤组织基因表达谱并从中筛选抗 菌肽相关基因, 采用新一代 Illumina 高通量测序平台对大弹涂鱼皮肤组织进行了转录组测序遥 利用 Trinity 软 件从头组装, 从测得的

78 608

366 条双端测序读长(paired-end read)共计

6 GB 的序列数据中获得

119 848 条高 质量的蛋白质编码基因(unigene)遥 经公共数据库序列检索和比对, 发现

7 个unigene 编码的多肽与已知的鱼类

5 大家族的抗菌肽高度同源, 即茁- 防御素 (茁-defensin)尧hepcidin尧NK-lysin尧piscidin 和肝表达抗菌肽 -2 (liver-ex- pressed antimicrobial peptide-2, LEAP-2)遥 最后, 对上述抗菌肽相关 unigene 开展了组织表达差异分析尧序列比对 及进化树分析遥 研究结果为进一步了解大弹涂鱼适应两栖生活的免疫防御机制和利用新鉴定的抗菌肽开发新 型抗菌药物奠定了基础遥 关键词: 大弹涂鱼;

皮肤;

转录组;

抗菌肽;

序列分析 中图分类号: Q71 文献标识码: A 文章编号: 1007-7847(2018)01-0026-10 Transcriptome of Boleophthalmus pectinirostris Skin and Analysis of Antimicrobial Peptide Related Unigenes ZHANG Yu-xia1 , SHI Ge1 , WANG Ri-xin2 , GONG Yan-bin1 , FAN Mei-hua1 , LIAO Zhi1* (1. Laboratory of Marine Biology Protein Engineering, Marine Science and Technical College, Zhejiang Ocean University, Zhoushan 316022, Zhejiang, China;

2. Marine Science College, Ningbo University, Ningbo 315211, Zhejiang, China) Abstract: Antimicrobial peptides (AMPs) are employed by fish as the innate defense peptides to protect against the invasion of foreign microorganisms and are important lead molecules for the development of novel drugs. Boleophthalmus pectinirostris is a special fish that can live in amphibious environment;

and the mucus from its skin is speculated to be rich in AMPs and has great significance to its immune defense and adaptation to the amphibious life. For understanding the gene expression patterns of B. pectinirostris skin and screening for antibacterial peptide related genes, the transcriptome analysis of the adult B. pectinirostris skin was per原formed by Illumina sequencing. A total of

78 608

366 paired-end reads amounting to

6 GB of sequence data were generated and further de novo assembled with Trinity software, yielding a total of

119 848 high-quality skin unigenes. When these unigenes were annotated in the available databases,

7 unigenes were found to en原code peptides homologous with the reported fish AMPs of