Replication of individual DNA molecules under electronic control using a protein nanopore.
Replication of individual DNA molecules under electronic control using a protein nanopore.
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使用蛋白质纳米孔在电子控制下复制单个 DNA 分子。
DOI：
10.1038/nnano.2010.177
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发表时间：
2010-11
影响因子：
38.3
通讯作者：
中科院分区：
文献类型：
--
作者：
纳米孔可以通过监测离子电流来分析DNA，因为单个链被捕获，并在施加电压的情况下以单一文件顺序通过孔。在这里，我们展示了单个DNA模板的连续复制可以通过保持在α-溶血素纳米孔口的DNA聚合酶来实现。复制在主体阶段被阻止，只有在DNA被纳米孔捕获后才开始复制。我们使用这种方法，结合主动电压控制，观察了噬菌体T7DNA聚合酶(T7DNAP)和DNA聚合酶I(KF)的Klenow片段催化的DNA复制。T7DNAP在DNA模板上前进，在纳米孔上施加80 mV的负载，并在毫秒级的时间尺度上测量了数百个串联的单个DNA分子的单核苷酸添加。当该酶以80 mV的外加电压放置在纳米孔口上时，没有观察到KF的复制。在受控的电压反转时，观察到KF的顺序核苷酸加成。
Nanopores can be used to analyse DNA by monitoring ion currents as individual strands are captured and driven through the pore in single file order by an applied voltage. Here we show that serial replication of individual DNA templates can be achieved by DNA polymerases held at the α-hemolysin nanopore orifice. Replication is blocked in the bulk phase, and is initiated only after the DNA is captured by the nanopore. We used this method, in concert with active voltage control, to observe DNA replication catalyzed by bacteriophage T7 DNA polymerase (T7DNAP) and by the Klenow fragment of DNA polymerase I (KF). T7DNAP advanced on a DNA template against an 80 mV load applied across the nanopore, and single nucleotide additions were measured on the millisecond time scale for hundreds of individual DNA molecules in series. Replication by KF was not observed when this enzyme was held atop the nanopore orifice at 80 mV applied potential. Sequential nucleotide additions by KF were observed upon controlled voltage reversals.
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影响因子：
15
作者：
Hurt N;Wang H;Akeson M;Lieberman KR
通讯作者：
Lieberman KR
影响因子：
15
作者：
Cockroft, Scott L.;Chu, John;Ghadiri, M. Reza
通讯作者：
Ghadiri, M. Reza
DOI：
10.1073/pnas.81.11.3297
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
ASSELINE, U;DELARUE, M;HELENE, C
通讯作者：
HELENE, C
影响因子：
5.6
作者：
Astatke, M;Grindley, NDF;Joyce, CM
通讯作者：
Joyce, CM
DOI：
10.1073/pnas.93.24.13770
发表时间：
1996-11-26
影响因子：
11.1
作者：
Kasianowicz, JJ;Brandin, E;Deamer, DW
通讯作者：
Deamer, DW