适合蜂王浆蛋白质组的双向电泳技术  

A Two-dimensional Gel Electrophoresis Protocol for Proteomic Study of Royal Jelly

作　　者：赵方圆[1,2] 吴亚君[1] 韩建勋[1] 陈颖[1] 葛毅强[2,3] 

机构地区：[1]中国检验检疫科学研究院,北京100123 [2]中国农业大学食品科学与营养工程学院,北京100083 [3]中国农村技术开发中心,北京100045

出　　处：《食品科学》2013年第9期14-18,共5页Food Science

基　　金：国家"863"计划项目(2011AA1008047)

摘　　要：为建立适用于蜂王浆蛋白质组研究的双向电泳技术,对其中的IPG胶条的选择、蛋白质上样量、等电聚焦时间及SDS-PAGE凝胶浓度等关键步骤进行探索和优化。结果显示:蜂王浆蛋白质pH值主要分布在pH 5～8范围内,当上样量为150μg,聚焦时间达到32000V.h的情况下能充分地聚焦,第二向使用10%的梯度胶时获得的双向电泳图谱最佳,在优化后的蜂王浆双向电泳图谱上可检测到62±4蛋白点,重复性高,分辨率较好。优化后的双向电泳技术体系适合于蜂王浆全蛋白质的双向电泳分析。In order to establish a two-dimensional gel electrophoresis（2-DE） protocol for proteomic study of royal jelly（RJ）,IPG strip with appropriate pH range,optimal loading amount for protein,focusing time of IEF,and separating gel concentration was used.The results showed that the best 2-DE image of RJ proteins using IPG strip with pH 5—8,protein loading amount of 150 μg,focusing time of 32000 V.h,and separating gel concentration of 10% was achieved.Moreover,62 ± 4 protein spots were detected on the gel with high repeatability and resolution.The 2-DE system after optimization is suitable for the analysis of RJ proteins,which provides theoretical basis for the proteomics of RJ.

关 键 词：蜂王浆 蛋白质组 双向电泳 优化 

分 类 号：TS201.6[轻工技术与工程—食品科学]