应用双向电泳和免疫印迹技术分析软叶针葵花粉变应原  

Analysis on the total allergens of phoenix roebelenii pollen with two dimensional electrophoresis followed by immunoblotting

作　　者：刘硕[1] 孟光[1] 郑琳麟 李春林[1] 谢伟伟[1] 龙绮[1] 

机构地区：[1]海口市人民医院耳鼻咽喉头颈外科,海口570208 [2]海口市人民医院中心实验室

出　　处：《临床耳鼻咽喉头颈外科杂志》2016年第11期878-880,共3页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：国家自然科学基金项目(No:81160129);海南省自然科学基金资助项目(No:30871;811145);海南省国际科技合作重点计划项目(No:2012-GH010);海口市重点科技计划项目[No:海科立2007-0080号;海科工信立(2012)72号];海南省卫生厅科研项目(No:琼卫2007-79)

摘　　要：目的：采用双向电泳和免疫印迹技术对软叶针葵花粉提取液蛋白质组分进行分析,寻找软叶针葵花粉的主要变应原。方法：提取软叶针葵花粉变应原,应用双向电泳（2-DE）结合免疫印迹技术,获得相应的电泳图谱和免疫印迹图谱。结果：软叶针葵花粉蛋白提取液经双向电泳后,用考马斯亮蓝染色,得到的电泳图显示约有601个主要蛋白斑点。分子量（Mr）分布约20～130kD;等电点（pI）主要集中在4.0～8.0。免疫印迹分析结果显示,阳性组与对照组的差异蛋白点数约为19个。结论：双向电泳联合免疫印迹技术对软叶针葵花粉提取液蛋白质组分分析,可寻找到其主要变应原,并可进一步用于变应原致敏组分的研究。Objective：To investigate allergen extract and to seek the main allergens of phoenix roebelenii pollen by the technology of two dimensional electrophoresis（2-DE）in protein analysis and immunoblotting.Method：Phoenix roebelenii pollen allergen extract was prepared with routine method,2-DE combined with immunoblotting assay（Western Blotting）,laser imagescanning,2-DE gel analysis software were used to analyzing its protein components.Result：About 601 soluble proteins spots were revealed in coomassie-stained gels.Most of the proteins had a molecular weight（Mr）of 20 to 130kD,and an isoelectric point（pI）value of 4.0to 8.0.Immunoblotting of 2-DE were showed and there were 19 specific antigen spots,compared with the controls.Conclusion：2-DE is a good method in protein analysis of ephemeroptera allergen extract,and it is useful in seeking main allergens and further research for allergic components.

关 键 词：软叶针葵花粉 变应原 双向电泳 免疫印迹 

分 类 号：R756.21[医药卫生—皮肤病学与性病学]