Vernet T, Dignard D, Thomas D Y
Gene. 1987;52(2-3):225-33. doi: 10.1016/0378-1119(87)90049-7.
The construction and characterization of a family of yeast expression vectors is described. They have the following features: plasmid replication and selection (ApR) in Escherichia coli, packaging of single-stranded (ss) DNA upon infection of E. coli with a filamentous helper phage, replication in Saccharomyces cerevisiae based on the 2 mu plasmid origin of replication (ori), selection in yeast by complementation of LEU2 (pVT-L series, size 6.3 kb) or URA3 gene (pVT-U series, size 6.9 kb) and seven unique restriction sites for cloning within an 'expression cassette' which includes the promoter and 3' sequence of the ADH1 gene. The multiple cloning site as well as the ori and intergenic region of the phage f1 have been cloned in two orientations for convenient gene cloning and ssDNA strand selection. As a result any of these eight vectors can be chosen for cloning, expressing genes in yeast, sequencing and mutagenesis without the need for recloning into specialized vectors.
本文描述了一系列酵母表达载体的构建及特性。它们具有以下特点：在大肠杆菌中进行质粒复制和筛选（氨苄青霉素抗性），丝状辅助噬菌体感染大肠杆菌后进行单链（ss）DNA包装，基于2μm质粒复制起点（ori）在酿酒酵母中复制，通过LEU2基因互补在酵母中筛选（pVT-L系列，大小6.3 kb）或URA3基因（pVT-U系列，大小6.9 kb），以及在一个“表达盒”内有七个用于克隆的独特限制酶位点，该表达盒包括ADH1基因的启动子和3'序列。噬菌体f1的多克隆位点以及ori和基因间区域已以两种方向进行克隆，便于基因克隆和单链DNA链选择。因此，可选择这八种载体中的任何一种进行克隆、在酵母中表达基因、测序和诱变，而无需重新克隆到专门的载体中。
