Discrimination of G-Quadruplexes from Duplex and Single-Stranded DNAs with Fluorescence and Energy-Transfer Fluorescence Spectra of Crystal Violet
Discrimination of G-Quadruplexes from Duplex and Single-Stranded DNAs with Fluorescence and Energy-Transfer Fluorescence Spectra of Crystal Violet
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用结晶紫的荧光和能量转移荧光光谱区分 G-四链体与双链体和单链 DNA
DOI：
10.1002/chem.200801441
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发表时间：
2009-01-01
影响因子：
4.3
通讯作者：
Shen, Han-Xi
中科院分区：
文献类型：
--
作者：
Kong, De-Ming;Ma, Yong-E.;Shen, Han-Xi
具有形成g -四重结构的潜力的富含g的核酸序列在生物学上重要的区域是常见的。这些序列大多与它们的互补链一起存在。因此，开发一种灵敏的生物传感器来区分g -四重结构和双工结构，并确定四重结构对双工结构的竞争能力受到了人们的广泛关注。在这项工作中。两种三苯基甲烷染料（孔雀石绿（MG）和结晶紫（CV））与g -四相、双相的相互作用。利用荧光光谱和能量转移荧光光谱对单链dna进行了研究。利用CV的荧光光谱或CV和MG的能量转移荧光光谱可以很好地区分四链和双链或单链dna。此外，利用能量转移荧光滴定法测定了CV与g-四聚体的结合化学计量比。通过荧光滴定具有富c互补链的g -四联体- cv复合物，可以测量在存在互补序列的情况下参与g -四联体结构的富g寡核苷酸的比例。本研究可能提供一种简单的方法来区分四重体和双链或单链dna，并在存在互补的富c序列时测量g -四重体百分比。
G-rich nucleic acid sequences with the potential to form G-quadruplex structures are common in biologically important regions. Most of these sequences are present with their complementary strands. so the development of a sensitive biosensor to distinguish G-quadruplex and duplex structure and to determine the competitive ability of quadruplex to duplex structures has received a great deal of attention. In this work. the interactions between two triphenylmethane dyes (malachite green (MG) and crystal violet (CV)) and G-quadruplex, duplex. or single-stranded DNAs were studied by fluorescence spectroscopy and energy-transfer fluorescence spectroscopy. Good discrimination between quadruplexes and duplex or single-stranded DNAs can be achieved by using the fluorescence spectrum of CV or the energy-transfer fluorescence spectra of CV and MG. In addition, by using energy-transfer fluorescence titrations of CV with G-quadruplexes, the binding-stoichiometry ratios of CV to G-quadruplexes call be determined. By using the fluorescence titrations of G-quadruplex-CV complexes with C-rich complementary strands, the fraction of G-rich oligonucleotide that engages in G-quadruplex structures in the presence of the complementary sequence can be measured. This Study may provide a simple method for discrimination between quadruplexes and duplex or single-stranded DNAs and for measuring G-quadruplex Percentages in the presence of the complementary C-rich sequences.