杂色曲霉菌变应原分离、鉴定及纯化  

Isolation,identification and partial purification of specific allergens from Aspergillus versicolor

作　　者：喻海琼[1,2] 周一平[1] 胡川[2] 李金生[2] 

机构地区：[1]深圳市福田区人民医院呼吸内科,广东518033 [2]南昌大学医学院

出　　处：《中国公共卫生》2011年第5期661-662,共2页Chinese Journal of Public Health

基　　金：深圳市科技计划项目(200902138)

摘　　要：目的对杂色曲霉菌变应原蛋白进行分离、鉴定与纯化。方法提取杂色曲霉菌蛋白质,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离,用免疫印迹(western-blot)鉴定变应原成分,用western-blot法鉴定出了6种变应原,通过离子交换和凝胶过滤层析纯化变应原。结果碳酸氢盐-盐水提取液(Coca’s液)共提取出23条蛋白质条带,分子量为17、20和62 kD的蛋白质为杂色曲霉菌主要变应原,而分子量为22、35和76kD的蛋白质为次要变应原;通过各种层析纯化变应原,得到含有分子量为20kD主要变应原和35kD的次要变应原的组分。结论鉴定了杂色曲霉菌的主要和次要变应原并部分纯化出其变应原组分,为变态反应疾病的诊断和治疗提供依据。Objective To isolate,identify and partially purify the allergens from fungal（Aspergillus versicolor）.Methods Proteins from Aspergillus versicolor were isolated and seperated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE）,and then identified by western-blot.The allergens of Aspergillus versicolor were consequently purified by ion exchange chromatography and gel chromatography.Results More than 20 kinds of allergen protein were separated from the crude extract of the fungus by SDS-PAGE and 6 allergens were identified by western-blot.The allergens with molecular weight（MW） of 17kD,20kD and 62kD had higher binding rate.The allergens with molecular weight of 22 kD,35 kDa,76 kDa were minor allergens.The fraction containing the major allergen with MW of 20kD and the minor allergen with MW of 35kD were purified.Conclusion The study identified major and minor allergens of Aspergillus versicolor and partially purified the allergens,which may provide a theoretic basis for allergy diagnosis and immunotherapy.

关 键 词：杂色曲霉菌 变应原 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE) 免疫印迹(western-blot) 离子交换层析 凝胶过滤层析 

分 类 号：Q786[生物学—分子生物学]