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YIpDCE1——一种用于酵母中双组成型表达的整合质粒。

YIpDCE1 - an integrating plasmid for dual constitutive expression in yeast.

作者信息

Stearman R, Dancis A, Klausner R D

机构信息

Cell Biology, Metabolism Branch, National Institute of Child Health, Human Development, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, USA.

出版信息

Gene. 1998 Jun 8;212(2):197-202. doi: 10.1016/s0378-1119(98)00179-6.

Abstract

YIpDCE1 (Dual Constitutive Expression), a novel Saccharomyces cerevisiae integrating plasmid, constitutively expresses two genes under the control of separate phosphoglycerol kinase promoters. YIpDCE1 contains the complete ADE2 gene which can be used as a marker for selecting integrants at mutant ade2 loci commonly present in laboratory yeast strains. The YIpDCE1 plasmid can be inserted into the ade2-101 locus of the HF7c strain used in two hybrid screens. Thus it could be useful for analysis of two hybrid interactions that occur in the context of additional protein components (e.g. modifying enzymes such as kinases or phosphatases, or multimeric complexes consisting of three or four distinct protein components). YIpDCE1 has been used to create strains simultaneously overexpressing the permease (FTR1) and oxidase (FET3) components of the yeast high-affinity iron uptake system. This confers constitutive high-affinity iron uptake on the transformed strains, bypassing the normal regulatory mechanisms.

摘要

YIpDCE1(双组成型表达)是一种新型的酿酒酵母整合质粒,在单独的磷酸甘油激酶启动子控制下组成型表达两个基因。YIpDCE1包含完整的ADE2基因,该基因可作为在实验室酵母菌株中常见的突变ade2位点选择整合体的标记。YIpDCE1质粒可插入用于双杂交筛选的HF7c菌株的ade2 - 101位点。因此,它对于分析在其他蛋白质成分(例如激酶或磷酸酶等修饰酶,或由三个或四个不同蛋白质成分组成的多聚体复合物)背景下发生的双杂交相互作用可能有用。YIpDCE1已被用于构建同时过表达酵母高亲和力铁摄取系统的通透酶(FTR1)和氧化酶(FET3)成分的菌株。这赋予转化菌株组成型高亲和力铁摄取能力,绕过了正常的调节机制。

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