摘要： 对酿酒酵母嗜杀质粒的提取、电泳检测及纯化方法进行了研究。以改进的Fried和Fink方法在不预破细胞壁的情况下,巯基乙醇在碱性条件(pH9. 3)预处理后,直接以SDS-苯酚抽提完整细胞分离到了嗜杀质粒(dsRNA) 。在此基础上, 以电泳透析方法获得了纯化的M2和L双链RNA,琼脂糖凝胶电泳测定分子大小分别为1.5kb和4.0kb。
Abstract:The method of extraction and purification of the double-stranded RNA killer plasmids from Saccharomyces cerevisiae was studied.With the improved method of Fried and Fink we directly extracted the intact cells with SDS-phenol after pre-treating the cells with ß-mercaptoethanol under analkli condition.The result was detected and confirmed by the gel electrophoresis.On this basis we acquired the purified M2 and L-dsRNA by means of electrophoretic dialysis,their molecular lengths being 1.5kb and 4.0kb respectively.