Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy
Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy
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DOI：
10.1093/nar/gkh572
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发表时间：
2004-04-01
影响因子：
14.9
通讯作者：
Knoll, W
中科院分区：
文献类型：
--
作者：
Tawa, K;Knoll, W
除了一个错配的碱基对(MM1)外，两个由互补碱基对组成的单链DNA可以通过分子识别形成双链DNA。这种类型的双工不如MM0形成的双工稳定。为了更好地了解传感器(芯片)表面杂交和解离过程的物理机制，我们用表面等离子体荧光光谱研究了MM1杂交的动力学。在5‘端标记荧光分子Cy5并与表面附着的探针DNA杂交的靶DNA链可以被表面等离子体共振模的强光场激发。发射的荧光可以高灵敏度地被检测到。双链的亲和力取决于化学性质，即G-G、G-T等，以及错配碱基对沿15聚双链的位置。
Two single-stranded DNAs consisting of complementary base pairs except for one mismatching base pair (MM1) can form double-stranded DNA by molecular recognition. This type of duplex is not as stable as that formed by MM0. In order to add to a better understanding of the physical mechanism of the hybridization and dissociation processes at sensor (chip) surfaces, we studied the kinetics of the MM1 hybridization by surface plasmon fluorescence spectroscopy. Target DNA strands labelled with a fluorescent molecule Cy5 at the 5' end and hybridizing with the surface-attached probe DNA can be excited by the strong optical field of a surface plasmon resonance mode. The emitted fluorescence can be detected with high sensitivity. The affinity of a duplex was found to depend on the chemical nature, i.e. G-G, G-T etc., and on the position of the mismatching base pair along the 15mer duplex.