基于DIA技术的转IGF-I基因细毛羊皮肤组织蛋白组学分析  

Proteomic Analysis of Skin Tissue in IGF-I Transgenic Fine Wool Sheep(Ovis aries)Based on DIA Technology

作　　者：张译元[1] 郭延华[1] 唐红[1] 高磊[1] 王新华[1] 周平[1] 王立民[1] ZHANG Yi-Yuan;GUO Yan-Hua;TANG Hong;GAO Lei;WANG Xin-Hua;ZHOU Ping;WANG Li-Min(Research Institute of Animal Husbandry and Veterinary Medicine,Xinjiang Academy of Agricultural and Reclamation Sciences/State Key Laboratory for Sheep Genetic Improvement and Healthy Production,Shihezi 832000,China)

机构地区：[1]新疆农垦科学院畜牧兽医研究所/省部共建绵羊遗传改良与健康养殖国家重点实验室,石河子832000

出　　处：《农业生物技术学报》2021年第5期943-951,共9页Journal of Agricultural Biotechnology

基　　金：国家转基因重大专项(2016ZX08008-001);国家自然科学基金(31660341,31760307)。

摘　　要：转胰岛素样生长因子-I(insulin-like growth factor I,IGF-I)基因的中国美利奴细毛羊(Ovis aries)(以下简称转基因细毛羊),羊毛产量得到显著提高。为探明与羊毛产量相关的细毛羊皮肤蛋白的表达变化,本研究利用数据非依赖性采集(data-independent acquisition,DIA)蛋白组测序技术和生物信息学分析,对转基因细毛羊和野生型细毛羊的皮肤总蛋白进行分析,以期筛选获得与羊毛生长特性相关的特异性蛋白。首先应用DIA蛋白质组学和液相色谱串联质谱(liquid chromatography-tandem mass spectrometry,LCMS/MS)方法对转IGF-I基因细毛羊和野生型细毛羊皮肤组织进行蛋白质组测序,对测序数据进行归一化处理、差异表达蛋白分析、基因本体(Gene Ontology,GO)和KEGG富集分析以及候选蛋白质筛选。结果表明,共计筛选获得2914个蛋白,差异表达蛋白930个,其中374个上调表达、556个下调表达,与羊毛性状相关的差异表达蛋白有18个;GO分析表明,生物学过程、分子功能、细胞组成相关的注释分别有24、14和18个;KEGG通路分析显示,差异表达蛋白涉及42条信号通路,其中丝氨酸/苏氨酸激酶、分泌型糖蛋白、促性腺激素释放激素、肿瘤抑制蛋白、转化生长因子β等8条信号通路可能与羊毛生长发育存在相关性。本研究对揭示差异表达蛋白与羊毛性状的关系有促进作用,为探明羊毛性状的分子调控机理及分子标记育种提供参考依据。The wool yield of transgenic Chinese Merion fine wool sheep(Ovis aries)with insulin-like growth factor I(IGF-I)gene was significantly increased.In order to find out the change of skin protein expression related to wool production,data independent acquisition(DIA)proteome sequencing technology and bioinformatics analysis were used to analyze the total skin protein of transgenic sheep and wild-type sheep,which aimed to find specific proteins related to wool growth characteristics.Firstly,DIA proteomics method and liquid chromatography tandem mass spectrometry(LC-MS/MS)were used to sequence the tissues of IGFI transgenic sheep and wild type sheep.The sequencing data were normalized,differentially expressed proteins were analyzed,Gene Ontology(GO)and KEGG enrichment analysis and candidate protein screening were carried out.The results showed that 2914 proteins were obtained,including 930 differentially expressed proteins.Compared with wild type,374 proteins were up-regulated and 556 proteins were down-regulated,and 18 differentially expressed proteins were related to wool traits.GO analysis showed that there were 24,14 and 18 annotations to biological process,molecular function and cell component,respectively;KEGG pathway analysis showed that the differentially expression proteins were involved in 42 signaling pathways,including 8 signaling pathways such as serine/threonine kinase,secretory glycoprotein,gonadotropin releasing hormone,tumor suppressor protein,transforming growth factor-β,which was related to wool growth and development.This study is helpful to reveal the relationship between differential protein and wool traits and provides a reference for exploring the molecular mechanism of wool traits and molecular marker breeding.

关 键 词：转基因 胰岛素样生长因子-I(IGF-I) 细毛羊 皮肤组织 数据非依赖性采集(DIA) 蛋白质组学 

分 类 号：S811[农业科学—畜牧学]