MAPPING REPLICATION UNITS IN ANIMAL-CELLS
MAPPING REPLICATION UNITS IN ANIMAL-CELLS
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DOI：
10.1016/0092-8674(89)90329-2
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发表时间：
1989-06-16
期刊：
影响因子：
64.5
通讯作者：
CEDAR, H
中科院分区：
文献类型：
--
作者：
HANDELI, S;KLAR, A;CEDAR, H
已经开发了用于测定任何DNA片段的体内复制方向的一般方法。该技术允许扫描基因组区域以检测双向尾对尾复制，从而指示功能性起点的存在。根据这一标准，我们确定了中国仓鼠DHFR基因下游两个起始位点的大致位置。进一步的映射揭示了头对头复制的区域，表示复制终止的位置，从而定义了完整动物细胞复制子的标志。DHFR起源存在的遗传学证据是通过显示该区域在其通过转染整合到基因组中的其他位点后作为双向DNA合成起始点而获得的。为了展示这种方法的普遍适用性，我们研究了APRT域。复制映射连同使用的缺失突变体允许在一个遥远的上游位点的起源的鉴定。
A general approach for assaying the in vivo direction of replication for any DNA segment has been developed. This technique allows the scanning of genomic regions to detect bidirectional tail-to-tail replication, indicating the presence of a functional origin. By this criterion we identified the approximate positions of two origin sites downstream of the Chinese hamster DHFR gene. Further mapping revealed areas of head-to-head replication, signifying locations of replication termination and thus defining the landmarks of a complete animal cell replicon. Genetic proof for the existence of the DHFR origin was obtained by showing that this region serves as a bidirectional DNA synthesis initiation point following its integration into other sites in the genome by transfection. To show the general applicability of this methodology, we studied the APRT domain. Replication mapping together with the use of deletion mutants allowed the identification of an origin at a far-upstream locus.