银杏营养贮藏蛋白质的分离鉴定

Isolation and Identification of Vegetative Storage Protein of Ginkgo biloba

采用光学和电子显微镜技术、SDS-PAGE技术、间接免疫酶标光镜定位和胶体金免疫电镜定位技术对银杏枝条中的营养贮藏蛋白质(VSPs)进行分离鉴定,并对其细胞学特性及动态变化规律进行初步研究。研究结果:运用制备型SDS-聚丙烯酰胺凝胶电泳分离并制备银杏32和36ku2种营养贮藏蛋白质抗原,通过免疫新西兰大白兔成功获得银杏32和36ku2种营养贮藏蛋白质的多克隆抗体。银杏VSPs主要分布于落叶期当年生枝条的次生韧皮部细胞中,呈现出团块状、颗粒状和均一状3种形态。在新梢生长期,积累在枝条中的VSPs逐渐降解消失,表现出明显的季节变化。

The vegetative storage proteins（VSPs） in branches of Ginkgo biloba were isolated and identified by using light-and electron-microscope,SDS-PAGE,immunoenzyme optical-microscopy localization and immunogold electron-microscopy localization.The cytological characteristic and dynamic changes of VSPs were preliminarily studied.The results were as follows：The antigens of vegetative storage protein of 32 and 36 ku were separated and prepared by using SDS-polyacrylamide gel electrophoresis,antibodies of vegetative storage proteins of 32 and 36 ku were successfully achieved with immuniting New Zealand rabbits.VSPs were found mainly in the secondary phloem parenchyma cells of one-year-old twigs during deciduous stage and they were accumulated in vacuole in different forms such as nubbly,granular, and homogeneous under electron microscope.The protein inclusions of branches almost disappeared during new shoot development,with obviously seasonal change.