Elledge S J, Mulligan J T, Ramer S W, Spottswood M, Davis R W
Department of Biochemistry, Baylor College of Medicine, Houston, TX 77030.
Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1731-5. doi: 10.1073/pnas.88.5.1731.
This work describes a multifunctional phage lambda expression vector system, lambda YES, designed to facilitate gene isolation from eukaryotes by complementation of Escherichia coli and Saccharomyces cerevisiae mutations. lambda YES vectors have a selection for cDNA inserts using an oligo adaptor strategy and are capable of expressing genes in both E. coli and S. cerevisiae. They also allow conversion from phage lambda to plasmid clones by using the cre-lox site-specific recombination system, referred to here as automatic subcloning. A simple method has been developed for the conversion of any plasmid into a phage lambda cDNA cloning vector with automatic subcloning capability. cDNA libraries constructed in these vectors were used to isolate genes from humans and Arabidopsis thaliana by complementation of yeast and bacterial mutations, respectively.
本研究描述了一种多功能噬菌体λ表达载体系统——λYES，该系统旨在通过互补大肠杆菌和酿酒酵母的突变来促进从真核生物中分离基因。λYES载体采用寡核苷酸接头策略筛选cDNA插入片段，并且能够在大肠杆菌和酿酒酵母中表达基因。它们还可以通过使用cre-lox位点特异性重组系统（本文称为自动亚克隆）从噬菌体λ转化为质粒克隆。已经开发出一种简单的方法，可将任何质粒转化为具有自动亚克隆能力的噬菌体λ cDNA克隆载体。分别通过互补酵母和细菌突变，利用构建在这些载体中的cDNA文库从人和拟南芥中分离基因。
