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作为一种遗传系统的酿酒酵母的重要性日益增长,从酵母转化子中分离质粒成为一门很重要的技术。通常使用两种质粒分离方案:第一种,从酵母转化子中直接分离质粒。此法的问题是通过限制酶分析鉴定重组质粒时,有同时分离内部2—μm环状DNA以及染色体DNA的潜在干扰。第二种方案,从酵母转化子中分离出DNA,用于转化大肠杆菌细胞。随后质粒可从细菌中既高产又可高质量的通过多种方法分离出来。但以上二方案均不能完全令人满意,方法耗时多而得到的结
The importance of Saccharomyces cerevisiae, a genetic system, is growing and the isolation of plasmids from yeast transformants has become an important technique. Two plasmid isolation protocols are commonly used: first, plasmids are directly isolated from yeast transformants. The problem with this method is the potential interference of simultaneous separation of the internal 2-μm circular DNA and chromosomal DNA when the recombinant plasmid is identified by restriction enzyme analysis. In the second scenario, DNA is isolated from yeast transformants and used to transform E. coli cells. Plasmids can then be separated from the bacteria by both high yield and high quality by a variety of methods. However, the above two programs are not completely satisfactory, the method takes too much time and the resulting knot