点带石斑鱼过敏原分离、鉴定与纯化
Isolation,identification and purification of allergens in grouper (epinephelus malabaricus)
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摘要: 目的 对点带石斑鱼的主要过敏原进行分离、鉴定与纯化.方法 通过十二烷基硫酸钠-聚丙炳酰胺凝胶电泳 (SDS-PAGE)分离点带石斑鱼的蛋白质组份,采用免疫印迹 (Western-blotting)方法鉴定过敏原,通过离子交换层析对主要过敏原进行初步纯化.结果 SDS-PAGE结果显示,点带石斑鱼粗提液含有13条蛋白带,含量高的有8条,分子量分别为43,34,28,25,22,17,15,9kD;Western-blotting显示,对点带石斑鱼过敏患者的阳性混合血清能与其中4个蛋白条带起反应,分子量分别为34,28,24,22kD.离子交换层析可初步纯化出22kD的过敏原蛋白.结论 初步分离得到分子量为22kD的点带石斑鱼主要过敏原,为临床诊断和治疗鱼类过敏性疾病提供理论依据.Abstract: Objective To isolate,identify and purify the allergens in grouper.Methods The total protein of the grouper was isolated by SDS-PAGE.The allergens of grouper were identified by Western-blotting.To purify the allergens,ion exchange chromatography was employed.Results There were 13 protein bands visible on the SDS-PAGE gel.The allergens with molecular mass of 34kD,28kD,25kD and 22kD were identified by Western-blotting.The 22kD allergen of grouper was purified by ion exchange chromatography.Conclusion The major allergens of grouper was isolated and identified for further study of diagnosis and therapy of fish allergy.
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