School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, UK.
J Exp Bot. 2012 Oct;63(16):6035-43. doi: 10.1093/jxb/ers252. Epub 2012 Sep 17.
The commercial cultivation of genetically engineered (GE) crops in Europe has met with considerable consumer resistance, which has led to vigorous safety assessments including the measurement of substantial equivalence between the GE and parent lines. This necessitates the identification and quantification of significant changes to the metabolome and proteome in the GE crop. In this study, the quantitative proteomic analysis of tomato fruit from lines that have been transformed with the carotenogenic gene phytoene synthase-1 (Psy-1), in the sense and antisense orientations, in comparison with a non-transformed, parental line is described. Multidimensional protein identification technology (MudPIT), with tandem mass spectrometry, has been used to identify proteins, while quantification has been carried out with isobaric tags for relative and absolute quantification (iTRAQ). Fruit from the GE plants showed significant alterations to their proteomes compared with the parental line, especially those from the Psy-1 sense transformants. These results demonstrate that MudPIT and iTRAQ are suitable techniques for the verification of substantial equivalence of the proteome in GE crops.
欧洲对基因工程(GE)作物的商业化种植遭到了消费者的强烈抵制,这导致了激烈的安全性评估,包括对 GE 品系与亲本系之间实质性等同性的测量。这就需要对 GE 作物的代谢组和蛋白质组进行鉴定和定量分析。在这项研究中,描述了与非转化亲本系相比,用反义方向和有义方向转化的类胡萝卜素生物合成基因phytoene synthase-1(Psy-1)的番茄果实的定量蛋白质组学分析。多维蛋白质鉴定技术(MudPIT)与串联质谱一起被用于鉴定蛋白质,而相对和绝对定量的同量异位标记(iTRAQ)则用于定量。与亲本系相比,GE 植株的果实的蛋白质组发生了显著改变,特别是 Psy-1 有义转化体的果实。这些结果表明,MudPIT 和 iTRAQ 适用于验证 GE 作物蛋白质组实质性等同性。