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酵母中染色体增加和倍性增加突变体的分离与鉴定

Isolation and characterization of chromosome-gain and increase-in-ploidy mutants in yeast.

作者信息

Chan C S, Botstein D

机构信息

Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

出版信息

Genetics. 1993 Nov;135(3):677-91. doi: 10.1093/genetics/135.3.677.

Abstract

We have developed a colony papillation assay for monitoring the copy number of genetically marked chromosomes II and III in Saccharomyces cerevisiae. The unique feature of this assay is that it allows detection of a gain of the marked chromosomes even if there is a gain of the entire set of chromosomes (increase-in-ploidy). This assay was used to screen for chromosome-gain or increase-in-ploidy mutants. Five complementation groups have been defined for recessive mutations that confer an increase-in-ploidy (ipl) phenotype, which, in each case, cosegregates with a temperature-sensitive growth phenotype. Four new alleles of CDC31, which is required for spindle pole body duplication, were also recovered from this screen. Temperature-shift experiments with ipl1 cells show that they suffer severe nondisjunction at 37 degrees. Similar experiments with ipl2 cells show that they gain entire sets of chromosomes and become arrested as unbudded cells at 37 degrees. Molecular cloning and genetic mapping show that IPL1 is a newly identified gene, whereas IPL2 is allelic to BEM2, which is required for normal bud growth.

摘要

我们开发了一种菌落乳头化检测方法,用于监测酿酒酵母中遗传标记的II号和III号染色体的拷贝数。该检测方法的独特之处在于,即使整个染色体组发生增加(多倍体增加),它也能检测到标记染色体的增加。此检测方法用于筛选染色体增加或多倍体增加的突变体。已针对赋予多倍体增加(ipl)表型的隐性突变定义了五个互补组,在每种情况下,该表型都与温度敏感型生长表型共分离。还从该筛选中获得了纺锤体极体复制所需的CDC31的四个新等位基因。对ipl1细胞进行的温度转换实验表明,它们在37℃时会发生严重的染色体不分离。对ipl2细胞进行的类似实验表明,它们会获得完整的染色体组,并在37℃时作为未出芽的细胞停滞生长。分子克隆和遗传定位表明,IPL1是一个新鉴定的基因,而IPL2与正常芽生长所需的BEM2等位。

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