Direct selection procedure for the isolation of functional centromeric DNA.
Direct selection procedure for the isolation of functional centromeric DNA.
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用于分离功能性着丝粒 DNA 的直接选择程序。
DOI：
10.1073/pnas.78.6.3760
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发表时间：
1981
影响因子：
11.1
通讯作者：
Carbon,J
中科院分区：
文献类型：
--
作者：
Hsiao,CL;Carbon,J
通过直接选择维持稳定的质粒，已从酵母基因组文库中分离出几个质粒，每个质粒都含有一段能够赋予在酵母（酿酒酵母）中复制的ars1载体YRp7有丝分裂稳定性的DNA片段。有丝分裂稳定序列分为两类：（i）酵母“2微米”质粒的片段和（ii）含有酵母着丝粒DNA的DNA片段。第一类质粒通过减数分裂分离 4+:0-，而着丝粒质粒主要分离 2+:2-，作为典型的染色体。先前鉴定的来自染色体 II (CEN3) 和 XI (CEN11) 的着丝粒以及 CEN11 连锁基因 MET14 可以通过这种直接选择程序分离。此外，还获得了其他几种未鉴定的着丝粒 DNA。 CEN3 和 CEN11 DNA 之间没有可检测到的序列同源性，这些 DNA 也不与其他酵母着丝粒区域杂交。
Several plasmids, each containing a segment of DNA capable of conferring mitotic stability on the ars1 vector YRp7 replicating in yeast (Saccharomyces cerevisiae), have been isolated from a yeast genomic library by direct selection for stable plasmid maintenance. The mitotic-stabilizing sequences fall into two classes: (i) segments of the yeast "2-micrometers" plasmid and (ii) DNA segments containing yeast centromeric DNA. Plasmids of the first class segregate 4+:0- through meiosis whereas the centromere plasmids segregate predominantly 2+:2-, as typical chromosomes. Previously identified centromeres from chromosomes II (CEN3) and XI (CEN11), along with the CEN11-linked gene MET14, can be isolated by this direct selection procedure. In addition, several other unidentified centromere DNAs were obtained. There are no detectable sequence homologies between CEN3 and CEN11 DNAs nor do these DNAs hybridize to the other yeast centromere regions.