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应用iTRAQ结合2D LC-MS/MS技术分析草菇同核和异核菌丝蛋白质组
引用本文:刘靖宇,江玉姬,谢宝贵,陈炳智,廖伟,邓优锦,刘朋虎.应用iTRAQ结合2D LC-MS/MS技术分析草菇同核和异核菌丝蛋白质组[J].食用菌学报,2012,19(1):1-12,105.
作者姓名:刘靖宇  江玉姬  谢宝贵  陈炳智  廖伟  邓优锦  刘朋虎
作者单位:1. 福建农林大学菌物研究中心,福建福州350002;山西农业大学,食品科学与工程技术学院,山西太谷030801
2. 福建农林大学菌物研究中心,福建福州,350002
基金项目:国家现代农业产业技术体系建设专项资金资助
摘    要:首次利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术结合二维液相色谱串联质谱对草菇(Volvariella volvacea)同核和异核菌丝蛋白质组进行研究。采用QExactive质谱鉴定并经MASCOT软件搜库,对所有鉴定蛋白进行主成分分析(principal component analysis,PCA)、层次聚类(hierarchical clustering)分析和GO(gene ontology)注释,并进行生物信息学分析。结果共获得2335个不同肽段,鉴定到1039个蛋白,其中1030个蛋白具有定量信息。在同核菌丝中显著上调蛋白83个,下调蛋白97个,表明iTRAQ标记技术结合二维液相色谱串联质谱可有效地分离和鉴定草菇菌丝蛋白质组。该研究结果为更深入全面地研究草菇菌丝蛋白质组学奠定基础。

关 键 词:草菇  蛋白质组  iTRAQ  二维液相色谱  串联质谱

Analysis of the Proteomes in Volvariella volvacea Homokaryotic and Heterokaryotic Mycelia by iTRAQ-coupled 2-D LC-MS/MS
LIU Jingyu , JIANG Yuji , XIE Baogui , CHEN Bingzhi , LIAO Wei , DENG Youjin , LIU Penghu.Analysis of the Proteomes in Volvariella volvacea Homokaryotic and Heterokaryotic Mycelia by iTRAQ-coupled 2-D LC-MS/MS[J].Acta Edulis Fungi,2012,19(1):1-12,105.
Authors:LIU Jingyu  JIANG Yuji  XIE Baogui  CHEN Bingzhi  LIAO Wei  DENG Youjin  LIU Penghu
Institution:1(1Mycological Research Center of Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;2Institute of Food Science and Engineer,Shanxi Agricultural University,Taigu,Shanxi 030801,China)
Abstract:Protein expression in homokaryotic and heterokaryotic mycelia of Volvariella volvacea has been investigated using isobaric tags for relative and absolute quantification(iTRAQ)-coupled 2D LC-MS/MS(two-dimensional liquid chromatography tandem mass spectrometry).A total of 1039 proteins,including 2335 unique peptides,were identified using Q Exactive mass spectrometry and the MASCOT search engine,and 1030 of these had a quantitative dimension.Of the quantitatively different proteins,83 were significantly up-regulated and 97 down-regulated in homokaryotic,compared with heterokaryotic,mycelium.Data from principal component analysis,hierarchical clustering and gene ontology(GO) also revealed that iTRAQ-coupled 2D LC-MS/MS represented a highly effective method for isolating and identifying protein groups in V.volvacea mycelia,and would be useful in future analyses of the fungal proteome.
Keywords:Volvariella volvacea  proteome  isobaric tags for relative and absolute quantification(iTRAQ)  two-dimensional liquid chromatography tandem mass spectrometry(2D LC-MS/MS)
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