Brachmann C B, Davies A, Cost G J, Caputo E, Li J, Hieter P, Boeke J D
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Yeast. 1998 Jan 30;14(2):115-32. doi: 10.1002/(SICI)1097-0061(19980130)14:2<115::AID-YEA204>3.0.CO;2-2.
A set of yeast strains based on Saccharomyces cerevisiae S288C in which commonly used selectable marker genes are deleted by design based on the yeast genome sequence has been constructed and analysed. These strains minimize or eliminate the homology to the corresponding marker genes in commonly used vectors without significantly affecting adjacent gene expression. Because the homology between commonly used auxotrophic marker gene segments and genomic sequences has been largely or completely abolished, these strains will also reduce plasmid integration events which can interfere with a wide variety of molecular genetic applications. We also report the construction of new members of the pRS400 series of vectors, containing the kanMX, ADE2 and MET15 genes.
已经构建并分析了一组基于酿酒酵母S288C的酵母菌株,其中根据酵母基因组序列通过设计删除了常用的选择标记基因。这些菌株将与常用载体中相应标记基因的同源性降至最低或消除,而不会显著影响相邻基因的表达。由于常用营养缺陷型标记基因片段与基因组序列之间的同源性已被大大降低或完全消除,这些菌株还将减少可能干扰多种分子遗传学应用的质粒整合事件。我们还报告了含有kanMX、ADE2和MET15基因的pRS400系列载体新成员的构建。