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作 者:杨慧[1] 刘志刚[1] 韩庆国[1] 侯穗波 梁桂珍
机构地区:[1]深圳大学生命科学院,广东深圳518060 [2]深圳市人民医院,广东深圳518020
出 处:《免疫学杂志》2004年第2期120-123,共4页Immunological Journal
基 金:国家自然科学基金资助项目 (30 0 70 70 2 )
摘 要:目的 对艾蒿、青蒿花粉变应原进行分离、鉴定。方法 采用不同的提取方式得到艾蒿、青蒿花粉的粗浸液 ,通过饱和 (NH4 ) 2 SO4 分级沉淀、聚丙烯酰胺凝胶电泳 (SDS PAGE)分离蛋白质组分 ,并用凝胶成像系统测定各组分的相对分子质量 (Mr) ;采用Western blotting鉴定 2种花粉的主要及次要变应原。结果 艾蒿、青蒿花粉分别分离到二十和十多种蛋白质组分。其中艾蒿花粉的组分中有 9种蛋白能与患者血清中蒿属花粉特异性IgE结合 ,Mr 为 6 2 0 0 0、4 30 0 0、380 0 0的蛋白条带的结合率最高。青蒿花粉的组分中有 11种蛋白能与患者血清中蒿属花粉特异性IgE结合 ,Mr 为 4 30 0 0、380 0 0的蛋白条带结合率最高。结论 艾蒿花粉的主要变应原Mr 分别为 6 2 0 0 0、4 30 0 0和 380 0 0 ,青蒿花粉的主要变应原Mr 分别为 4 30 0 0和 380 0 0 ;2种花粉变应原组分存在很大相似性 。Objective To isolate and identify the Artemisia argyi and Artemisia apiacea pollen. Methods Artemisia argyi and Artemisia apiacea pollen extract was precipitated by saturated ammonium sulfate and then electrophoresed by SDS-polyacrylamide gel (SDS-PAGE). The relative molecular weight (M r) of each protein band was determined by Gel Media System. We identified the primary and secondary allergen proteins by Western-blotting. Results We isolated more than twenty protein bands from Artemisia argyi pollen extract. The results showed that 9 of the protein bands can be combined with the specific IgE from sera of known Artemisia pollen allergic patients. The protein bands whose M r are 62 000,43 000 and 38 000 have the highest binding capacity. Similarly, we isolated more than ten protein bands from Artemisia apiacea pollen extract. The results showed that 11 of the protein bands can be combined with the specific IgE from sera of known Artemisia pollen allergic patients. The protein bands whose M r are 43 000,38 000 have the highest binding capacity. Conclusion The primary allergens of Artemisia argyi include the allergen proteins whose M r are 62 000,43 000 and 38 000,and the primary allergens of Artemisia apiacea include the allergen proteins whose M r are 43 000 and 38 000. Pollen from both species shares many allergen components. Unique allergen protein bands in each pollen allergens were also identified respectively.
关 键 词:青蒿花粉 艾蒿花粉 SDS-PAGE Western-blotting
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