采用一种能同时提取RNA、DNA和Protein的试剂(RDP试剂)进行日本囊对虾性腺总蛋白质的制备,并通过双向电泳(2-DE)技术与传统的裂解液法比较提取效果.RDP法所得2-DE图谱清晰,条纹少,蛋白质点数多,而裂解液法存在横向和纵向拖尾,其碱性端蛋白质点大量缺失,说明RDP法能有效去除蛋白质样品中盐离子、脂类等干扰成分.结果不仅为后续的日本囊对虾性腺蛋白质组学研究提供了技术保障,也为其他甲壳类动物性腺总蛋白质的提取提供了借鉴.
The gonad total protein of penaeid shrimp Marsupenaeus japonicus was prepared by a special reagent used to extract DNA, RNA and protein simultaneously (RDP reagent). The extraction efficiency was compared with traditional lysis solution method by two-dimensional eleetrophoresis (2-DE). The 2-DE map of RDP was clear with less striations and more protein spots, while that of lysis solution method had horizontal and vertical striations with decrease of protein spots at the cathodic side. These results indicated that RDP method could remove salt and lipid from gonad sample effectively. The finding from the study provided not on- ly technical support for proteomies research of M. japonicus , but also useful reference for gonad total protein extraction of other crustacean.