Marsoni Milena, Bracale Marcella, Espen Luca, Prinsi Bhakti, Negri Alfredo S, Vannini Candida
Dipartimento Ambiente-Salute-Sicurezza, Università dell'Insubria, via J.H. Dunant 3, 21100 Varese, Italy.
Plant Cell Rep. 2008 Feb;27(2):347-56. doi: 10.1007/s00299-007-0438-0. Epub 2007 Sep 15.
Two dimensional gel electrophoresis coupled to mass spectrometry has been used to study the somatic embryogenesis in Vitis vinifera, by comparing embryogenic and non embryogenic calluses of the Thompson seedless cv. More than 1,000 spots were reproducibly resolved in colloidal Coomassie brilliant blue stained gels over a pI nonlinear range of 3-10 in the first dimension and using homogeneous 12.5% polyacrylamide gels in the second dimension. The expression pattern of 35 spots differed significantly between the two samples. These spots were processed by mass spectrometry analysis and the protein identity was assigned by using both the non-redundant protein and EST databases. Several responsive proteins, some already known to be involved in the somatic embryogenesis process while others, for the first time put into relation with this process, have been described. Moreover, they have been subdivided in functional categories, and their putative role is discussed in terms of their relevance in the somatic embryogenesis process.
二维凝胶电泳结合质谱分析法已被用于研究葡萄的体细胞胚胎发生,通过比较汤普森无核葡萄品种的胚性愈伤组织和非胚性愈伤组织。在第一维中,在3-10的非线性pH范围内,用考马斯亮蓝胶体染色凝胶可重复分辨出1000多个斑点,第二维使用均匀的12.5%聚丙烯酰胺凝胶。两个样品之间35个斑点的表达模式有显著差异。这些斑点通过质谱分析进行处理,并通过使用非冗余蛋白质和EST数据库来确定蛋白质身份。已经描述了几种响应蛋白,其中一些已知参与体细胞胚胎发生过程,而其他一些则首次与该过程建立了联系。此外,它们已被细分为功能类别,并根据它们在体细胞胚胎发生过程中的相关性讨论了它们的推定作用。