GENETIC-ANALYSIS OF THE MITOTIC TRANSMISSION OF MINICHROMOSOMES
GENETIC-ANALYSIS OF THE MITOTIC TRANSMISSION OF MINICHROMOSOMES
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DOI：
10.1016/0092-8674(85)90153-9
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发表时间：
1985-01-01
期刊：
影响因子：
64.5
通讯作者：
HARTWELL, LH
中科院分区：
文献类型：
--
作者：
KOSHLAND, D;KENT, JC;HARTWELL, LH
酿酒酵母微小染色体有丝分裂传递的保真度是通过一种新的目测方法来监测的，该方法检测单个有丝分裂中的质粒拷贝数的变化。本实验用于研究含有酵母复制起点(ARS1)和着丝粒(CEN3)的有丝分裂传递。微染色体的不正确分离的比率比观察到的正常染色体高200倍。然而，微小染色体的复制受到严格的控制；它的过度复制不到一次/1000有丝分裂。该方法还可用于分离和鉴定ARS1和CEN3的突变。ARS1中的突变定义了其最佳活性所需的新结构域，CEN3中的突变表明元素II的完整性对着丝粒功能不是必不可少的。最后，ARS1和CEN3突变的表型分别与它们在复制和分离中的功能一致。
The fidelity of the mitotic transmission of minichromosomes in Saccharomyces cerevisiae is monitored by a novel visual assay that detects changes in plasmid copy number in individual mitotic divisions. This assay is used to investigate the mitotic transmission of a plasmid containing a putative yeast origin of replication (ARS1) and a centromere (CEN3). The rate of improper segregation for the minichromosome is 200-fold higher than observed for a normal chromosome. However, the replciation of the minichromosome is stringently controlled; it overreplicates less than once/1000 mitotic divisions. This assay is also used to isolate and characterize mutations in ARS1 and CEN3. The mutations in ARS1 define a new domain required for its optimal activity, and the mutations in CEN3 suggest that the integrity of element II is not essential for centromere function. Finally, the phenotypes of the mutations in ARS1 and CEN3 are consistent with their function in replication and segregation, respectively.