中国武汉地区葎草花粉变应原蛋白质组分的双向电泳分析

应用蛋白质双向电泳的分析技术对武汉地区生长的主要过敏原———葎草(Humulus scandensLour.)花粉的蛋白质组分进行了分析。应用三氯乙酸法提取葎草花粉总蛋白质,通过等电聚焦和第二向SDS-聚丙烯酰胺凝胶电泳分析获得了完整的葎草花粉全蛋白质图谱。应用专业分析软件(Im ageM aster 2D)对电泳图谱分析表明:通过等电聚焦和第二向SDS-PAGE电泳,有122个不同的蛋白质组分被检测出来,并进一步确定每个蛋白质相应的分子量、等电点和相对含量。研究中获得的高分辨率的双向电泳图谱是我国葎草花粉变应原蛋白质第一张完整的蛋白质图谱,将为今后葎草花粉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基础。

Analysis of Total Proteins in Pollen of Humulus scandens Lour. in Wuhan Region of China by Two-dimensional Electrophoresis

The total proteins in pollen of Humulus scandens Lour.,one of the most popular aeroallergen in China,were analyzed by two-dimensional electrophoresis in current study.The total proteins of the pollen 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.After analysis with software(ImageMaster 2D),122 different proteins were detected;their isoelectric point(pI),Molecular weight(MW) and relative volume of each protein in the pollen were also discovered.This is the first high-resolution,two-dimensional protein map of the pollen of H.scandens Lour.in China.Our finding has built a solid foundation for identification,characterization,gene cloning and standardization of allergenic proteins in the pollen of H.scandens Lour.in further.