DNA detection system using molecularly imprinted polymer as the gel matrix in electrophoresis
DNA detection system using molecularly imprinted polymer as the gel matrix in electrophoresis
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DOI：
10.1016/j.bios.2006.08.026
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发表时间：
2007-04-15
影响因子：
12.6
通讯作者：
Shimizu, Toshimi
中科院分区：
文献类型：
--
作者：
Ogiso, Masayo;Minoura, Norihiko;Shimizu, Toshimi
为了开发一种简单而廉价的DNA检测方法，我们制备了一种分子印迹聚合物（MIP）来识别特定的双链DNA （dsDNA）序列，并将其用于电泳凝胶基质。MIP凝胶具有许多结合位点，这些结合位点在大小、形状和目标dsDNA序列官能团的排列上是互补的。在MIP凝胶电泳（MIPGE）过程中，MIP凝胶中结合位点的捕获效应会阻碍目标dsDNA的迁移。通过观察DNA标准尺寸标记在聚丙烯酰胺凝胶和MIP凝胶中的迁移距离之间的线性关系的偏差，证实了这一点。非靶dsDNA的迁移距离则保持线性关系。此外，利用Ha-ras基因及其点突变体研究了该方法中dsDNA的序列选择性。除了A（.）从T到T碱基对替换，突变的dsDNA(例如，从A（。）从T到C。源自G（.）从C到TA)可以与目标（野生型）dsDNA区分开来。尽管A（.）有所改善。T（T(.)A）碱基对的区分仍然是需要的，本研究首次证明了用MIPs检测特定的dsDNA序列，因此，为MIPs的实际应用开辟了一个新的领域。(c) 2006 Elsevier B.V.版权所有
To develop a simple and inexpensive method for DNA detection, we prepared a molecularly imprinted polymer (MIP) for recognizing a specific double-stranded DNA (dsDNA) sequence and used it in an electrophoretic gel matrix. The MIP gel has many binding sites that are complementary in size, shape, and arrangement of functional groups of the target dsDNA sequence. During MIP gel electrophoresis (MIPGE), migration of the target dsDNA should be hindered by the capture effect of the binding sites in the MIP gel. This was confirmed by observation of deviations from the linear relationship between the migration distances of the DNA standard size markers in the polyacrylamide gel and those in the MIP gel. The migration distances of nontarget dsDNA maintained a linear relationship, however. In addition, the sequence selectivity of dsDNA in this method was investigated by using the Ha-ras gene and its point mutants. Except for A(.)T to T(.)A base pair substitution, mutant dsDNA (for example, substitution from A(.)T to C(.)G and from G(.)C to T(.)A) could be distinguished from the target (wild-type) dsDNA. Although some improvement in A(.)T (T(.)A) base pair distinction is still needed, this study is the first to demonstrate detection of a specific dsDNA sequence with MIPs and, as such, opens up a new realm for practical applications of MIPs. (c) 2006 Elsevier B.V. All rights reserved.