论文部分内容阅读
目的对短穗鱼尾葵花粉过敏原蛋白质组分进行双向电泳及其结果分析,构建短穗鱼尾葵花粉蛋白双向电泳图谱,明确短穗鱼尾葵花粉的过敏原蛋白组分的分布和构建蛋白质组学数据库。方法提取短穗鱼尾葵花粉的总蛋白,取上清液进行Bradford法定量总蛋白浓度,采用13 cm IPG胶条(p H 3~10)和12%的SDS-PAGE凝胶作为二项分离胶进行双向电泳(2D)分离,银染法分析其总蛋白组成,利用Image Master2D软件对2D胶进行检测和图像分析。结果双向电泳图谱共检测到516个短穗鱼尾葵花粉蛋白点,其蛋白相对分子质量在12~174 k D,主要在10~50 k D;等电点在p H 3~10,主要分布在p H 5~8。含量最多的蛋白质其相对分子质量和等电点分别为13 k D和p H 6.258 37。含量最低的蛋白质其相对分子质量和等电点分别为29 k D和p H 3.615 79。结论成功构建较为完整的短穗鱼尾葵花粉蛋白双向电泳图谱,为建立短穗鱼尾葵花粉蛋白质组学数据库及其过敏的诊断和治疗提供了依据。
Aim To analyze the protein components of pollen allergens from two populations and analyze the results. The two-dimensional gel electrophoresis profiles of pollen protein of Haliotis discus hannai were established, and the distribution and construction of allergen protein components Proteomics database. Methods The total protein was extracted from the pollen grains of Fritillaria cirrhosa, and the supernatant was used to determine the total protein concentration by Bradford method. The 13 cm IPG strips (p H 3 ~ 10) and 12% SDS-PAGE gel Glue was separated by two-dimensional electrophoresis (2D), the total protein composition was analyzed by silver staining and the 2D glue was detected and analyzed by Image Master2D software. Results Two hundred and seventy-two spots were detected in the two-dimensional gel electrophoresis. The relative molecular mass of the protein was 12 ~ 174 kD, mainly in the range of 10 ~ 50 kD; the isoelectric point was in p H 3 ~ 10, In p H 5 ~ 8. The highest content of protein relative molecular mass and isoelectric point were 13 k D and p H 6.258 37 respectively. The protein with the lowest content has a relative molecular mass and isoelectric point of 29 kD and pH3.61579, respectively. Conclusion The two-dimensional electrophoresis pattern of the complete polysaccharides of Pleurotus citrinopileatus was successfully constructed, which provided a basis for the establishment of the database and allergy diagnosis and treatment of pollen of Pleurotus.