AUTONOMOUS DNA-REPLICATION IN HUMAN-CELLS IS AFFECTED BY THE SIZE AND THE SOURCE OF THE DNA
AUTONOMOUS DNA-REPLICATION IN HUMAN-CELLS IS AFFECTED BY THE SIZE AND THE SOURCE OF THE DNA
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DOI：
10.1128/mcb.11.4.2263
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发表时间：
1991-04-01
影响因子：
5.3
通讯作者：
CALOS, MP
中科院分区：
文献类型：
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作者：
HEINZEL, SS;KRYSAN, PJ;CALOS, MP
我们以前开发了用于人类细胞中DNA自主复制的短期和长期测定。 本研究讨论了这些试验中的复制要求。 将62个大小为1至21 kb的随机人基因组片段克隆到原核载体中，并在短期试验中测试其复制能力。 我们发现复制强度和片段长度之间呈正相关，表明大尺寸有利于人类细胞中高效的自主复制。 所有大片段都能有效复制，这表明可以指导人类细胞中DNA复制起始的信号要么非常丰富，要么具有低程度的序列特异性。 在长期试验中获得了相似的结果。 我们还使用相同的检测方法在人体细胞中检测了一系列来自大肠杆菌染色体DNA的随机片段。 在短期和长期试验中，细菌片段支持复制的效率低于人类片段。 这一结果表明，虽然参与人类细胞复制的序列信号经常在人类DNA中发现，但它们在细菌DNA中并不常见。
We previously developed short-term and long-term assays for autonomous replication of DNA in human cells. This study addresses the requirements for replication in these assays. Sixty-two random human genomic fragments ranging in size from 1 to 21 kb were cloned in a prokaryotic vector and tested for their replication ability in the short-term assay. We found a positive correlation between replication strength and fragment length, indicating that large size is favored for efficient autonomous replication in human cells. All large fragments replicated efficiently, suggesting that signals which can direct the initiation of DNA replication in human cells are either very abundant or have a low degree of sequence specificity. Similar results were obtained in the long-term assay. We also used the same assays to test in human cells a random series of fragments derived from Escherichia coli chromosomal DNA. The bacterial fragments supported replication less efficiently than the human fragments in the short-term and long-term assays. This result suggests that while the sequence signals involved in replication in human cells are found frequently in human DNA, they are uncommon in bacterial DNA.