双向电泳分析宣威火腿蛋白质组方法的建立

食品科学 ›› 2012, Vol. 33 ›› Issue (23): 125-128.

双向电泳分析宣威火腿蛋白质组方法的建立

廖国周1，王桂瑛2，程志斌1，施忠芬2，曹锦轩3,*

1.云南农业大学食品科技学院 2.云南农业职业技术学院畜牧兽医系 3.宁波大学生命科学与生物工程学院

收稿日期:2011-10-07 修回日期:2012-10-23 出版日期:2012-12-15 发布日期:2012-12-12
通讯作者: 廖国周 E-mail:lgzwgy@126.com
基金资助:
云南省社会发展科技计划应用基础研究专项;云南省现代农业生猪产业技术体系专项

Proteomic Analysis of Xuanwei Ham by Two-Dimensional Gel Electrophoresis

Received:2011-10-07 Revised:2012-10-23 Online:2012-12-15 Published:2012-12-12
Contact: Guo-Zhou LIAO E-mail:lgzwgy@126.com

摘要/Abstract

摘要： 以宣威火腿为对象，采用三氯乙酸-丙酮沉淀法提取宣威火腿肌肉蛋白，对蛋白定量后，进行一向等电聚焦电泳和二向聚丙烯酰胺凝胶电泳，电泳后的凝胶以考马斯亮蓝染色，ImageMaster 2D Platinum软件分析凝胶上的蛋白质点，切取1个蛋白质点，经胰蛋白酶消化后使用基质辅助激光解析电离-串联飞行时间质谱(MALDI-TOF/TOF MS)获取肽质量指纹图谱，并用Mascot软件分析，以建立宣威火腿的蛋白质组学研究方法。结果表明：经双向电泳图谱分析所获得的宣威火腿蛋白点分布于pH4.0～7.0之间，共发现1749个蛋白点，蛋白点清晰，图谱分辨率较好；切取的蛋白质点经鉴定为Gamma-2-肌动蛋白，Mascot检索分值为129分(阈值为67分)。可见，成功建立双向电泳分析宣威火腿蛋白质组的方法。

关键词: 宣威火腿, 蛋白质组学, 双向电泳

Abstract: In the present study, a method for the proteomic analysis of Xuanwei ham was established using two-dimensional gel electrophoresis (2-DE). Muscle protein from Xuanwei ham was extracted with trichloroacetic acid-acetone, quantified, and analyzed by isoelectric focusing electrophoresis and SDS-PAGE. After electrophoresis, the gels were stained with coomassie brilliant blue, and the observed protein spots were analyzed using imageMaster 2D platinum software and one of them was selected and analyzed by matrix-assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF/TOF MS) after tripsin digestion. The obtained peptide mapping fingerprint was analyzed using Mascot software. A total of 1749 clear protein spots were observed in the 2-DE pattern of Xuanwei ham, which were mainly distributed over a pH range of 4.0－7.0. The tested protein spot was identified as gamma-2-actin protein and found to have a Mascot score of 129 (threshold was 67). These results demonstrate the successful establishment of a 2-DE method for the proteomic analysis of Xuanwei ham.

Key words: Xuanwei ham, proteomics, two-dimensional electrophoresis

中图分类号:

TS207.3

廖国周1，王桂瑛2，程志斌1，施忠芬2，曹锦轩3,*. 双向电泳分析宣威火腿蛋白质组方法的建立[J]. 食品科学, 2012, 33(23): 125-128.

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