AUTONOMOUS REPLICATION IN HUMAN-CELLS OF MULTIMERS OF SPECIFIC HUMAN AND BACTERIAL-DNA SEQUENCES
AUTONOMOUS REPLICATION IN HUMAN-CELLS OF MULTIMERS OF SPECIFIC HUMAN AND BACTERIAL-DNA SEQUENCES
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DOI：
10.1128/mcb.13.5.2688
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发表时间：
1993-05-01
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：
KRYSAN, PJ;SMITH, JG;CALOS, MP
使用特定的2，712-bp人类DNA序列和特定的2，557-bp大肠杆菌DNA序列的模块，我们创建了包含1至12个单一或嵌合序列组成模块的质粒，并在人类细胞中测试了它们的自主复制能力。我们发现，每代的复制效率随着人类模块的连续添加而增加，通过六个拷贝基本上达到100%。虽然单个拷贝的细菌模块具有可忽略的复制能力，但每代12个细菌模块的复制效率为66%。由人和细菌模块组成的嵌合体显示中间复制水平。我们还使用了二维凝胶电泳来物理地绘制复制在半人半大肠杆菌上开始的位置。coli质粒。我们的研究结果表明，人类细胞中的自主复制是由简单的序列特征刺激的，这些特征经常出现在人类DNA中，但在细菌DNA中更为罕见。
Using modules of a specific 2,712-bp human DNA sequence and a specific 2,557-bp Escherichia coli DNA sequence, we created plasmids containing between 1 and 12 modules of single or chimeric sequence composition and tested them in human cells for their autonomous replication ability. We found that replication efficiency per generation increased with successive addition of human modules, to essentially 100% by six copies. Although a single copy of the bacterial module had negligible replication ability, the replication efficiency per generation of 12 bacterial modules was 66%. Chimeras composed of human and bacterial modules displayed intermediate replication levels. We also used two-dimensional gel electrophoresis to physically map where replication initiated on a half human-half E. coli plasmid. Our results suggest that autonomous replication in human cells is stimulated by simple sequence features which occur frequently in human DNA but are more rare in bacterial DNA.