Vilhena Milca B, Franco Mônica R, Schmidt Daiana, Carvalho Giselle, Azevedo Ricardo A
Departamento de Genética 11, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, BR.
An Acad Bras Cienc. 2015 Sep;87(3):1853-63. doi: 10.1590/0001-3765201520150116. Epub 2015 Aug 25.
Proteomics is an outstanding area in science whose increasing application has advanced to distinct purposes. A crucial aspect to achieve a good proteome resolution is the establishment of a methodology that results in the best quality and wide range representation of total proteins. Another important aspect is that in many studies, limited amounts of tissue and total protein in the tissue to be studied are found, making difficult the analysis. In order to test different parameters, combinations using minimum amount of tissue with 4 protocols for protein extraction from tomato (Solanum lycopersicum L.) leaves and roots were evaluated with special attention to their capacity for removing interferents and achieving suitable resolution in bidimensional gel electrophoresis, as well as satisfactory protein yield. Evaluation of the extraction protocols revealed large protein yield differences obtained for each one. TCA/acetone was shown to be the most efficient protocol, which allowed detection of 211 spots for leaves and 336 for roots using 500 µg of leaf protein and 800 µg of root protein per gel.
蛋白质组学是科学领域中一个杰出的领域，其应用的不断增加已服务于不同的目的。实现良好蛋白质组分辨率的一个关键方面是建立一种能够产生最佳质量和广泛代表性的总蛋白的方法。另一个重要方面是，在许多研究中，发现待研究组织中的组织和总蛋白量有限，这使得分析变得困难。为了测试不同参数，评估了使用最少组织量并结合4种从番茄（Solanum lycopersicum L.）叶片和根中提取蛋白质的方案的组合，特别关注它们去除干扰物的能力以及在二维凝胶电泳中实现合适分辨率的能力，以及令人满意的蛋白质产量。对提取方案的评估显示，每种方案获得的蛋白质产量差异很大。三氯乙酸/丙酮被证明是最有效的方案，使用每块凝胶500μg叶片蛋白和800μg根蛋白时，该方案可检测到叶片中的211个斑点和根中的336个斑点。
