Moleculer Surgery of DNA Using Enzyme-Immobilized Particles
Moleculer Surgery of DNA Using Enzyme-Immobilized Particles
复制标题
使用酶固定颗粒进行 DNA 分子手术
DOI：
10.1541/ieejsmas.116.196
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发表时间：
1996
影响因子：
--
通讯作者：
N. Shimamoto
中科院分区：
文献类型：
--
作者：
M. Washizu;Takatoki Yamamoto;O. Kurosawa;Seiichi Suzuki;N. Shimamoto
本文提出，并通过实验证明，一种新的方法的“分子手术”的DNA，其中酶标记的颗粒或探针被压对DNA，这是固定在基板上具有已知的构象和方向，使化学修饰在任意位置的DNA分子。该方法利用微加工电极产生的高强度高频电场（≥ 1 MV/m，≤ 1 MHz），通过静电取向效应将DNA拉伸成与电场平行的直线构象，并通过介电泳将DNA定位在基底上。一旦DNA被这样拉伸和定位，就可以访问任何一个对齐分子的任意位置。用DNA切割酶DNaseII标记直径为2.8μm的乳胶颗粒，然后进行激光操作并与DNA接触。观察到DNA在接触点处被瞬间切割。这一结果被认为是空间分辨生化修饰的第一个证明。
This paper proposes, and experimentally demonstrates, a novel method of 'molecular surgery' of DNA, where a enzyme-labeled particle or probe is pressed against DNA, which is immobilized on the substrate with known conformation and orientation, to make chemical modifications at arbitrary positions of the DNA molecule. In the method presented in this paper, DNA is subjected to high-intensity high-frequency electric field (≥1MV/m, ≈1MHz) created in micro-fabricated electrodes, so that it is 1) stretched to a straight conformation parallel to the field by the electrostatic orientation effect, and 2) positioned onto a substrate by the dielectrophoresis. Once DNA is thus stretch-and-positioned, one has access to arbitrary position of any one of aligned molecules. A latex particle, 2.8μm in diameter, is labeled with DNA-cutting enzyme, DNaseII, and is laser-manipulated and contacted with DNA. It is observed that DNA is instantaneously cut at the contact point. This result is considered to be the first demonstration of space-resolved biochemical modification.