Wait Robin, Miller Ingrid, Eberini Ivano, Cairoli Fausto, Veronesi Cristina, Battocchio Massimiliano, Gemeiner Manfred, Gianazza Elisabetta
Kennedy Institute of Rheumatology Division, Faculty of Medicine, Imperial College of Science, Technology and Medicine, London, UK.
Electrophoresis. 2002 Sep;23(19):3418-27. doi: 10.1002/1522-2683(200210)23:19<3418::AID-ELPS3418>3.0.CO;2-7.
A reference map for Bos taurus serum was obtained using proteomic tools: 21 proteins, plus several serum albumin fragments, have been identified in 47 spots. One of the major acute-phase reactants, haptoglobin, was also detected in a pathological serum. A number of technical problems had to be solved. (i) Spot resolution in two-dimensional electrophoresis (2-DE) is not easily optimized, as several proteins have similar molecular mass; different polyacrylamide concentration gradients were used for the analysis of various size ranges. (ii) Identification of proteins through mass spectrometry (MS) procedures is also difficult as the genome of Bos taurus is incompletely characterized. The program FASTS proved particularly useful, since it allows simultaneous searching of several unordered sequence fragments, which may be individually too short to provide a statistically valid match using BLAST.
在47个斑点中鉴定出了21种蛋白质以及几种血清白蛋白片段。主要急性期反应物之一的触珠蛋白也在一份病理血清中被检测到。有一些技术问题必须解决。(i) 二维电泳（2-DE）中的斑点分辨率不易优化，因为几种蛋白质具有相似的分子量；针对不同大小范围的分析使用了不同的聚丙烯酰胺浓度梯度。(ii) 通过质谱（MS）程序鉴定蛋白质也很困难，因为牛的基因组特征尚未完全明确。事实证明，FASTS程序特别有用，因为它允许同时搜索几个无序的序列片段，这些片段单独来看可能太短，无法使用BLAST提供统计学上有效的匹配。
