溴化乙锭标记DNA电化学探针的研究

以乙基-(3-二甲基丙基)碳化二亚胺盐酸盐(EDC)为偶联活化剂,将电化学活性物质溴化乙锭(Ethidiumbromide,EB)成功地标记在人工合成的含有21个碱基的寡聚DNA片段上,制备成EB标记DNA探针;用电化学方法将待测样品DNA片段固定在石墨电极表面,在一定的温度、pH值和离子强度条件下与EB标记DNA探针进行杂交反应,从而对靶序列DNA片段进行识别和测定.此外,还讨论了该探针的电化学性质、荧光光谱、待测DNA片段在石墨电极表面的电化学固定、DNA链碱基长度对EB标记DNA电化学探针的影响以及探针的选择性、重现性和寿命,结果令人满意.     

DNA探针在羧基聚苯乙烯微珠表面的微阵列研究

报道在１－乙基－３－３－（３－二甲基氨丙基）－碳化二亚胺（ＥＤＣ）偶链剂存在下,５’－ＮＨ_２单末端和５’－Ｔｅｘ／３’－ＮＨ_２双末端修饰ＤＮＡ探针在１０μｍ 羧基功能微珠表面的微阵列特性．研究表明,ＤＮＡ探针在微珠表面的微阵列能力决定于溶液介质的ｐＨ,ＤＮＡ探针浓度和微珠总表面积．优化条件下, ２０ ｍｅｒ单链ＤＮＡ探针在微珠表面微阵列的最小分子间距约为１４ ｎｍ,而２０ ｍｅｒ双链ＤＮＡ分子微阵列的最小分子间距约为２７ ｎｍ．如果ＤＮＡ探针序列长度由 ２０增加到 ３５ ｍｅｒ,探针在微珠表面的微阵列密度降低．机理研究表明,ＤＮＡ探针在羧基功能微珠表面的微阵列方式是探针分子近平行于微珠表面．     

基于纳米金探针和基因芯片的DNA检测新方法

运用荧光纳米金探针和基因芯片杂交建立一种新的DNA检测方法. 荧光纳米金探针表面标记有两种DNA探针: 一种为带有Cy5荧光分子的信号探针BP1, 起信号放大作用; 另一种为与靶DNA一部分互补的检测探针P532, 两种探针比例为5∶1. 当靶DNA存在时, 芯片上捕捉探针(与靶DNA的另一部分互补)通过碱基互补配对结合靶DNA, 将靶DNA固定于芯片上; 荧光纳米金探针通过检测探针与靶DNA及芯片结合, 在芯片上形成“三明治”复合结构, 最后通过检测信号探针上荧光分子的信号强度来确定靶DNA的量. 新方法检测灵敏度高, 可以检测浓度为1 pmol/L的靶DNA, 操作简单, 检测时间短. 通过改进纳米金探针的标记和优化杂交条件, 可进一步提高核酸检测的灵敏度, 这将在核酸检测方面具有重要的应用价值.     

自猝灭荧光探针法测定DNA

随着现代分子生物学和分子生物技术的发展，体内重要的遗传物质──脱氧核糖核酸（DNA）越来越引起人们的重视，对于DNA探针的研究也日渐深入．DNA探针作为研究DNA的有力工具，在90年代取得了长足的进展，新的靶扩增技术（PCR）已日臻完善，发光标记探针也已经与放射性同位素标记具有同等重要的地位，而且已应用于DNA杂交研究、疾病诊断和食品微生物检测等方面[1~5]．但是，研究开发新的、灵敏度高的DNA探针仍然是十分迫切和有意义的工作．本文报道了一种新的基于水杨酸、邻氨基苯甲酸荧光自碎灭的DNA探针技术，这对筛选灵敏度高…     

一种新型发光探针与DNA相互作用的研究

本文以稳态荧光光谱、紫外-可见吸收光谱、荧光偏振、热变性、阴离子猝灭等手段,研究了一种具有强电荷转移能力的化合物2,7-二[(N-乙基咔唑-3-基)丙烯-1-酮基]芴与DNA的相互作用。研究结果表明,2,7-二[(N-乙基咔唑-3-基)丙烯-1-酮基)芴与DNA的作用方式是混合模式,以嵌插作用为主,同时存在沟槽相互作用,其咔唑基团可能插入到DNA的碱基对之间,结合常数K为8 123.48mol/L,结合位点n为0.71。该发光探针灵敏度高,结合稳定。     

铽（Ⅲ）探针法研究一些金属离子与DNA的作用

本文首次选用Ｔｂ（Ⅲ）作为荧光探针，研究了ｃｄ（Ⅱ）、Ｚｎ（Ⅱ）、Ｃａ（Ⅱ）、Ｍｇ（Ⅱ）与ＤＮＡ反应的结合部位。结果表明：Ｃｄ（Ⅱ）、Ｚｎ（Ⅱ）在ＤＮＡ上有两种结合部位，即ＤＮＡ双螺旋外侧的磷酸基和内侧的碱基鸟漂呤上的Ｎ－Ｔ。而ｃａ（Ⅱ）、Ｍｇ（Ⅱ）在ＤＮＡ上只有一个结合部位即磷酸基。说明Ｔｂ（Ⅲ）不仅可以作为ＤＮＡ构象及ＤＮＡ单链成分、不配对碱基残余的探针，而且可以用它探测一些金属离子与ＤＮＡ反应的结合部位。     

电化学DNA生物传感器的研究现状

对电化学DNA生物传感器研究的现状,主要对1996-2006年期间的工作作了评述。内容涉及此类生物传感器的研究及DNA修饰电极与小分子的相互作用,还对此领域的未来发展作了展望（引用文献49篇）。     

DNA探针技术及其在生物分析化学上的应用

DNA探针广泛应用于基因工程、分子生物学研究及医学检验等领域,它在生物分析化学上的意义日益受到人们重视。DNA 探针的化学标记分为放射性核素标记和非放射性标记两类。非放射性标记物,主要包括酶、荧光和化学发光试剂以及半抗原物质。检测标记物可以利用习见的分析化学手段。从 DNA 探针技术的发展前景看,化学工作者在该领域是可以大有作为的。     

CdTe纳米荧光探针的合成及其与DNA的相互作用

以巯基丙酸(RSH)为稳定剂,采用水相法合成了功能性CdTe纳米晶,并通过X射线衍射(XRD)和透射电镜(TEM)对其粒度和形貌进行表征。建立了一种以水溶性CdTe量子点作为荧光探针测定DNA的方法,当DNA浓度为0.2~40μmol.dm-3时,荧光强度与DNA浓度呈良好的线性关系,检测限为80nmol.dm-3,11次重复测定含有5.6μmol.dm-3的小牛胸ctDNA得到的相对标准偏差为3.4%。考察了CdTe量子点浓度、pH值、温度及作用时间等因素对DNA荧光强度的影响。研究发现CdTe纳米粒子与DNA之间存在强烈的相互作用,量子点的荧光猝灭与DNA浓度呈线性关系;作用机理研究表明,CdTe纳米粒子与DNA之间存在静电相互作用,且DNA对CdTe纳米粒子的猝灭为动态猝灭过程。     

A New CoII Complex as a Bulge-Specific Probe for DNA

Bulge cleavage of two or three bases occurs when a DNA substrate is specifically cleaved oxidatively by [Co^II(tfa)₂(happ)] (see picture). Hydrogen peroxide is necessary for the activation of this octahedral complex, which suggests that hydroxyl radicals are the reactive species. The complex has no significant reactivity towards the corresponding sequence in a single-stranded DNA region, and it exhibits only a low affinity towards double-stranded DNA. happ=macrocyclic ligand based on 1,10-phenanthroline, tfa=trifluoroacetate.     

G-rich sequence-specific recognition and scission of human genome by PNA/DNA hybrid G-quadruplex formation

Hole in one: A single peptide nucleic acid (PNA) effectively targets the G-rich region in double-stranded DNA through formation of a PNA/DNA hybrid G-quadruplex. Only one target site in the whole human genome was selectively cleaved by the hybrid G-quadruplex. Such site-selective scission of DNA is central to gene manipulation for molecular biology, biotechnology, and therapy.     

桑色素及其配合物与DNA作用的比较

采用光度法、粘度法和电化学方法考察了桑色素及其配合物和DNA的作用.并对桑色素合铜(Ⅱ)、桑色素合锌(Ⅱ)、桑色素合钴(Ⅱ)、桑色素与DNA作用的方式进行了比较.Cu(Ⅱ), Zn(Ⅱ), Co(Ⅱ)桑色素配合物与DNA作用时的荧光光谱性质类似,不同于配体.钴(Ⅱ)配合物的电化学行为与铜(Ⅱ)配合物相似,不同于锌(Ⅱ)配合物.三种配合物的存在,都可引起DNA的粘度增加.但钴(Ⅱ)配合物对EB-DNA复合物的荧光光谱影响很小,而铜(Ⅱ)、锌(Ⅱ)配合物对EB-DNA复合物的荧光光谱影响较大.实验结果表明,与DNA之间的作用,钴(Ⅱ)配合物弱于铜(Ⅱ)、锌(Ⅱ)配合物.这可能是钴(Ⅱ)配合物的抗肿瘤活性低于与DNA嵌入结合的铜(Ⅱ)、锌(Ⅱ)配合物的原因之一.     

Berberrubine Phosphate: A Selective Fluorescent Probe for Quadruplex DNA

A phosphate-substituted, zwitterionic berberine derivative was synthesized and its binding properties with duplex DNA and G4-DNA were studied using photometric, fluorimetric and polarimetric titrations and thermal DNA denaturation experiments. The ligand binds with high affinity toward both DNA forms (K_b = 2–7 × 10⁵ M⁻¹) and induces a slight stabilization of G4-DNA toward thermally induced unfolding, mostly pronounced for the telomeric quadruplex 22AG. The ligand likely binds by aggregation and intercalation with ct DNA and by terminal stacking with G4-DNA. Thus, this compound represents one of the rare examples of phosphate-substituted DNA binders. In an aqueous solution, the title compound has a very weak fluorescence intensity (Φ_fl < 0.01) that increases significantly upon binding to G4-DNA (Φ_fl = 0.01). In contrast, the association with duplex DNA was not accompanied by such a strong fluorescence light-up effect (Φ_fl < 0.01). These different fluorimetric responses upon binding to particular DNA forms are proposed to be caused by the different binding modes and may be used for the selective fluorimetric detection of G4-DNA.     

DNA与Rt、VB2相互作用的电化学性质

芦丁(Rt)是存在于多种植物体中的一种多羟基黄酮化合物,是某些中草药的有效成分。本文在前文基础上着重对芦丁、VB2在玻碳电极上的电化学性质、二者与DNA相互作用的机理等方面的差异进行比较,这对维生素类药物与生物大分子相互作用的研究与应用具有重要意义。     

A Peptide Nucleic Acid (PNA)‐DNA Ferrocenyl Intercalator for Electrochemical Sensing

A ferrocenyl intercalator was investigated to develop an electrochemical DNA biosensor employing a peptide nucleic acid (PNA) sequence as capture probe. After hybridization with single strand DNA sequence, a naphthalene diimide intercalator bearing ferrocene moieties (FND) was introduced to bind with the PNA‐DNA duplex and the electrochemical signal of the ferrocene molecules was used to monitor the DNA recognition. Electrochemical impedance spectroscopy was used to characterize the different modification steps. Differential pulse voltammetry was employed to evaluate the electrochemical signal of the FND intercalator related to its interaction with the complementary PNA‐DNA hybrid. The ferrocene oxidation peaks were utilised for the target DNA quantification. The developed biosensor demonstrated a good linear dependence of FND oxidation peak on DNA concentration in the range 1 fM to 100 nM of target DNA, with a low detection limit of 11.68 fM. Selectivity tests were also investigated with a non‐complementary DNA sequence, indicating that the FND intercalator exhibits a selective response to the target PNA‐DNA duplex.     

Real Time Electrochemical Monitoring of DNA/PNA Dissociation by Melting Curve Analysis

An immobilization‐free electrochemical method is reported for real‐time monitoring of the DNA hybrid dissociation between a ferrocene labeled peptide nucleic acid (PNA) and a fully‐complementary or single‐base‐mismatched DNA. This method takes advantages of electrostatic charge characteristics and interactions among the neutrally charged PNA, the negatively charged DNA and the negatively charged electrode surface made of indium tin oxide (ITO). When a ferrocene labeled PNA (Fc‐PNA) sequence is hybridized to a complementary DNA strand, electrostatic repulsion between the negatively charged PNA/DNA hybrid and the negative ITO surface retards the diffusion of the electroactive Fc to the electrode, resulting in a much reduced electrochemical signal. On the other hand, when the Fc‐PNA is dissociated from the hybrid at elevated temperatures, the neutrally charged Fc‐PNA easily diffuses to the electrode with an enhanced electrochemical signal. Therefore, an electrochemical melting curve of the Fc‐PNA/DNA hybrid can be obtained by measuring the Fc signal with the increasing temperature. This strategy allows monitoring of the dissociation of the DNA hybrid in real time, which might lead to a simple detection method for single nucleotide polymorphism (SNP) analysis.     

寡聚酰胺为探针的电化学DNA生物传感器

DNA分子中的碱基对可以长程传递电荷, DNA分子中的碱基π堆积结构为电荷的长程传递提供了良好的通道. 电荷在DNA分子中的传递受碱基序列的影响, 利用这种性质可以构建DNA碱基错配检测的电化学传感器. 寡聚酰胺能和DNA以小沟绑定方式高亲和力地结合, 并且具有序列识别功能, 本文以带有硝基官能团的寡聚酰胺分子为电化学探针, 设计了电化学DNA生物传感器. 结果显示, 寡聚酰胺与DNA修饰电极作用后, 电化学响应显著增强, 并且可以作为检测DNA碱基错配的电化学探针分子.     

用于细胞核成像的2,7-BHVC双光子荧光探针

分别合成了2,7-双(1-羟乙基-4-乙烯基吡啶) 咔唑碘盐(2,7-BHVC)与3,6-BHVC, 进行了荧光性能和细胞核成像测试, 结果表明, 2,7位的Ф×δ是3,6位的2.8~3.6倍, 染色实验也证实了2,7-BHVC有特定成像细胞核的能力.