A FAMILY OF VERSATILE CENTROMERIC VECTORS DESIGNED FOR USE IN THE SECTORING-SHUFFLE MUTAGENESIS ASSAY IN SACCHAROMYCES-CEREVISIAE
A FAMILY OF VERSATILE CENTROMERIC VECTORS DESIGNED FOR USE IN THE SECTORING-SHUFFLE MUTAGENESIS ASSAY IN SACCHAROMYCES-CEREVISIAE
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DOI：
10.1016/0378-1119(88)90202-8
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发表时间：
1988-10-30
期刊：
影响因子：
3.5
通讯作者：
DAVIS, RW
中科院分区：
文献类型：
--
作者：
ELLEDGE, SJ;DAVIS, RW
一个简单的分析称为扇区洗牌被开发来监测克隆到酵母着丝粒质粒的必需基因的突变状态。该测定的本质是在不存在该基因的条件性温敏或冷敏等位基因的情况下，为必需基因创建条件性表型，即菌落颜色分区。这允许通过观察克隆的必需基因对试验菌株的扇形表型的影响来快速确定其突变状态。在这项工作的过程中，我们开发了一个家庭的20大肠杆菌-酵母穿梭载体，pUN质粒，含有ARS 1 CEN 4和各种选择标记，以及SUP 11基因，可以作为一个颜色标记在适当的背景。这些载体是紧凑的，并已非常有用的扇区洗牌测定和一般的基因分析。本文介绍了这些载体，扇区洗牌和几个应用程序的扇区表型。
A simple assay called the sectoring shuffle was developed to monitor the mutational state of essential genes cloned into yeast centromeric plasmids. The essence of this assay is the creation of a conditional phenotype, colony color sectoring, for an essential gene in the absence of conditional thermosensitive or cold-sensitive alleles of that gene. This allows the quick determination of the mutational state of a cloned essential gene by observing its effect on the sectoring phenotype of the tester strain. During the course of this work we developed a family of 20 Escherichia coli-yeast shuttle vectors, pUN plasmids, containing ARS1 CEN4 and a variety of selectable markers as well as the SUP11 gene which can act as a color marker in the proper background. These vectors are compact and have been very useful for the sectoring-shuffle assay and for gene analysis in general. This paper describes these vectors, the sectoring shuffle and several applications of sectoring phenotypes.