食入性过敏原杏仁蛋白组分的双向电泳分析

Analysis of total proteins in the seed of almond (Prunus dulcis) by two-dimensional electrophoresis

目的 :应用蛋白质双向电泳的分析技术 ,对一种主要的食入性过敏原杏仁果实的蛋白质组分进行分析。方法 :通过三氯乙酸法提取杏仁果实总蛋白质 ,通过等电聚焦和第 2向SDS PAGE分析获得完整的杏仁果实全蛋白质的图谱 ,并应用相应分析软件 (ImageMaster 2D)对电泳图谱进行分析。结果 :通过等电聚焦和第 2向SDS PAGE ,有 188个不同的蛋白质组分被检测出来。其中大约 2 8%的蛋白质等电点 (pI)在 4 .5～ 5 .5之间 ,大约 6 2 %的蛋白质的相对分子质量 (Mr)在 (2 0～ 2 5 )× 10 3 之间。结论 :所获得的高分辨率的双向电泳图谱 ,是我国杏仁果实蛋白质第一张完整的蛋白质图谱 ,为今后杏仁果实中致敏蛋白质的检测 ,分离、基因克隆和变应原的标准化奠定了基础。

AIM: To analyse the total proteins in the seeds of almond (Prunus dulcis), one of the popular ingestent allergens in China, by two-dimensional electrophoresis. METHODS: The total proteins of the seeds were extracted by trichloracetic acid (TCA) method, and then separated by isoelectric focusing as first dimension and SDS-PAGE as the second dimension. The spots of proteins were visualized by staining with Coomassie Brilliant Blue R-250. RESULTS: After analysis with software (ImageMaster 2D), 188 different proteins were detected. The isoelectric points (pI) for approximately 28% of total proteins were between 4.5-5.5, and the relative molecular mass (M r) of approximately 62% total proteins were between (20-25)×103. CONCLUSION: This was the first high-resolution, two-dimensional protein map of the seed of almond (Prunus dulcis) in China. Our finding has laid a solid foundation for further identification, characterization, gene cloning and standardization of allergenic proteins in the seed of almond (Prunus dulcis).