一种简便的适用于酵母双杂交系统的酵母质粒提取方法  

A Simple Method for Isolation of Yeast Plasmid Used for Yeast Two-Hybrid System

作　　者：苑丽娜[1] 王定和[2] 王永杰[3] 郭佳[2] 单卫星[2,4] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [2]西北农林科技大学植物保护学院,陕西杨凌712100 [3]河南省郸城县第一高级中学,河南郸城477150 [4]陕西省农业分子生物学重点实验室,陕西杨凌712100

出　　处：《生物技术通讯》2011年第6期864-866,共3页Letters in Biotechnology

基　　金：高等学校学科创新引智计划(B07049);现代农业产业技术体系(nycytx-15)

摘　　要：目的:建立一种适用于酵母双杂交系统的简便快捷的酵母质粒提取方法。方法:以酿酒酵母为供试材料,用玻璃珠振荡法破除酵母细胞壁,提取酵母总DNA,最后通过电转化大肠杆菌DH10B获得目的质粒。结果:粗提得到的质粒可直接转化DH10B,作为模板用于PCR分析及酵母双杂交后续的序列分析等,大大降低了工作量。结论:该方法简便快捷,经济实用,降低了成本,提高了效率,可以作为一种实验室酵母质粒提取方法。Objective： To establish a fast,stable and economical isolation method of plasmid from yeast for the yeast two-hybrid,a glass beads-based simple method for isolation of plasmid DNA from yeast Saccharomyces cerevisiae was developed.Methods： Yeast cells were mixed with glass beads and the target plasmids released by vortexing,preceipitated and transformed into E.coli DH10B by electroporation.Results： Agarose gel electrophoresis showed that the quality of the extracted plasmid in the crude yeast plasmid DNA was enough for PCR of candidate genes identified from yeast two-hybrid screening.Conclusion： The described method is simple,fast and low-cost,allowing facilitated large scale isolation and analysis of candidate genes involved in the use of yeast systems.

关 键 词：玻璃珠法质粒提取 酵母质粒 酵母双杂交 

分 类 号：Q78[生物学—分子生物学]