摘要
报道了一种从团头鲂卵壳中提取卵壳蛋白的有效方法。该提取液经SDS PAGE电泳鉴定后 ,证明有三条主要的蛋白带 ,其分子量分别为 6 4KDa,5 6KDa和 5 2KDa ,借助于制备型SDS PAGE电泳和电洗脱分离纯化技术 ,获得了三种高纯度的卵壳蛋白。
This study deals with an effective procedure for extraction and purification of zona radiata proteins (Zrps) from eggshells of Bluntnose black bream (Megalobrama amblycephala Yih). To avoid harding resulting from parthenogenic activation, unfertilized eggs were immediately immersed in a large volume of ice cold buffer A. After homogenization and washing, the eggshells were frozen dry and solubilized in buffer B. The SDS-PAGE electrophoretic migration pattern shows the solubilized Zrps consiste mainly of three protein bands with the apparent molecular weight of 64KDa, 56KDa and 52KDa. To purify these three Zrps, the solubilized eggshell proteins were subjected to preparative SDS-PAGE. The protein bands were visualized using an ice-cold KCl solution. The opaque bands were excised from the gel and extracted using extraction buffer with electro-elution. SDS-PAGE pattern of purified Zrps shows highly purified Zrps were acquired and sufficient for further studies with immunoassay.
出处
《水生生物学报》
CAS
CSCD
北大核心
2003年第2期132-135,共4页
Acta Hydrobiologica Sinica
基金
国家自然科学基金资助项目 (No .2 0 1770 32 )
武汉市晨光计划项目 (No .995 0 0 40 96G)
国家 86 3项目 (No.2 0 0 1AA6 40 6 0 1)资助
关键词
团头鲂
卵壳蛋白
提取
电洗脱
纯化
Bluntnose black bream
Zona radiata proteins
Extraction
Electro-elution
Purification
