利用Mascot程序与家蚕EST数据对质谱数据进行鉴定.pdfVIP

中国农业科学 2010,43(12):2561-2569 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2010.12.022 利用Mascot程序与家蚕EST数据对质谱数据进行鉴定 刘晓勇，刘海军，陈克平，姚 勤 （江苏大学生命科学研究院，江苏镇江 212013 ） 摘要：【目的】研究和检验利用Mascot程序与家蚕EST数据解析质谱分析的结果，鉴定目标蛋白的方法。【方 法】将家蚕5龄起蚕中肠总蛋白进行二维电泳分离，并运用质谱对80个蛋白点进行肽指纹图谱分析，利用Mascot 程序和经自主编制软件Transeq翻译的EST 序列数据库对所得质谱数据进行鉴定。【结果】鉴定出其中41个蛋白， 并利用 EST 电子克隆在其余 39 个蛋白中得到 19 个蛋白的全长或较长 cDNA。【结论】本文提出的利用家蚕 EST 数 据来解析家蚕蛋白质谱结果的方法，能够鉴定仅利用公共数据不能鉴定的蛋白，并能鉴别基因可变剪接后产生蛋 白异形体的问题。 关键词：家蚕；中肠；质谱；表达序列标签；肽指纹图谱 Identification of Mass Spectral Data Using Mascot Program and Silkworm EST Data LIU Xiao-yong, LIU Hai-jun, CHEN Ke-ping, YAO Qin (Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, Jiangsu) Abstract: 【Objective 】 Methodology for identification of silkworm (Bombyx mori) protein using mass spectra data, Mascot program and EST data was evaluated. 【Method 】 Total protein of the fifth instar silkworm midgut was separated by two-dimensional electrophoresis, and the peptide fingerprintings of 80 protein spots were analyzed using mass spectrometry. Those mass spectral data were identified using Mascot program and silkworm EST database translated by a software called Transeq. 【Result 】 Forty-one proteins were identified and the full-length or longer cDNA of 19 proteins among other 39 proteins were obtained using the EST in silico cloning. 【Conclusion 】 The methodology could be used to identify proteins using mass spectral data with silkworm EST data, as well as alternatively spliced variants. Key words: silkworm; midgut; mass spectra; expressed sequence tag; peptide mass fingerprinting 质[5] 。测定肽混合物质量数最有效的质