Systematic Plasmid Libraries for Genetic Analysis in Yeast

随机基因组DNA文库的发展是一项重要的技术突破，使任何基因都能够物理分离。随机文库继续用于基因筛选和克隆基因，但它们存在的问题是，并不是基因组的所有部分都存在，而且并不是所有部分的水平都相同。本项目的目标是通过构建酿酒酵母基因组的系统文库来克服这些问题。在实现这一目标方面取得了重大进展，但物质和功能上的差距依然存在。这些空白将被填补，来自整个文库的插入将被转移到另一个载体，创建一对匹配的系统化文库，用于低水平表达和每个酵母基因的过度表达。所得到的文库将使高效和全面的过度表达筛选成为可能，并有助于通过质粒互补来克隆基因。

The development of random genomic DNA libraries was an important technological breakthrough that enabled the physical isolation of any gene. Random libraries continue to be used in genetic screens and for cloning genes, but they suffer from the problems that not all portions of the genome are present, and that not all portions are present at equal levels. The goal of this project is to overcome these problems by constructing systematic libraries of the Saccharomyces cerevisiae genome. Significant progress has been made towards this goal, but physical and functional gaps remain. These gaps will be filled, and the inserts from the entire library will be transferred to another vector, creating a matching pair of systematic libraries for low level expression and for overexpression of every yeast gene. The resulting libraries will enable efficient and comprehensive overexpression screens and facilitate cloning of genes by plasmid complementation.