Bonneaud N, Ozier-Kalogeropoulos O, Li G Y, Labouesse M, Minvielle-Sebastia L, Lacroute F
Centre de Génétique Moléculaire du C.N.R.S., Laboratoire propre associé à l'Université Pierre et Marie Curie, Gif sur Yvette, France.
Yeast. 1991 Aug-Sep;7(6):609-15. doi: 10.1002/yea.320070609.
We describe a set of replicative, integrative and single-stranded shuttle vectors constructed from the pUC19 plasmid that we use routinely in our experiments. They bear a yeast selectable marker: URA3, TRP1 or LEU2. Replicative vectors carrying different yeast replication origins have been constructed in order to have plasmids based on the same construction with a high or low copy number per cell and with different mitotic stabilities. All the vectors are small in size, provide a high yield in Escherichia coli and efficiently transform Saccharomyces cerevisiae. These plasmids have many of the unique sites of the pUC19 multicloning region and many of them allow for the screening of plasmids with an insert by alpha-complementation. The nucleotide sequence of each of them is completely known.
我们描述了一组从pUC19质粒构建的复制型、整合型和单链穿梭载体,这些载体我们在实验中经常使用。它们带有一个酵母选择标记:URA3、TRP1或LEU2。为了获得基于相同构建、每个细胞具有高拷贝数或低拷贝数以及不同有丝分裂稳定性的质粒,已构建了携带不同酵母复制起点的复制型载体。所有载体尺寸小,在大肠杆菌中产量高,并能高效转化酿酒酵母。这些质粒具有pUC19多克隆区域的许多独特位点,其中许多允许通过α互补筛选带有插入片段的质粒。它们每一个的核苷酸序列都是完全已知的。