Carbazole tricationic salt: A novel potential two-photon fluorescent DNA probe for nucleic imaging of cells

A new carbazole tricationic salt, 4,4′-(1E,1′E)-2,2′-(9-(2-(1-(2-hydroxyethyl)pyridinium-4-yl)ethyl)-9H-carbazole-3,6-diyl) bis(ethane- 2,1-diyl) bis(1-(2-hydroxyethyl)pyridinium) iodide (THEPC) was synthesized. Photophysical experiments have shown that THEPC has large two-photon excited fluorescence action cross-sections (33 GM in the presence of DNA), which ranks THEPC as a good biological fluorophore. The results from electronic absorption, circle dichroism and single-/two-photon fluorescence emis-sion spectra suggest that THEPC can strongly bind to DNA, with an intrinsic binding constant of 5.79 × 106 L mol–1. THEPC has better photostability under one- or two-photon excitation conditions. Finally, the staining photos from two-photon fluorescence microscopy (TPM) show that THEPC can exclusively label the nucleus with high contrast and without image distortion. These re-markable properties and optimized imaging ability make THEPC an attractive DNA probe in TPM.     

4-苄氧基苯酚与鲱鱼精DNA相互作用的研究

在pH=7.4的Tris-HCl缓冲溶液中,采用荧光光谱法、黏度法等分析方法研究了4-苄氧基苯酚(PBP)与鲱鱼精DNA的作用方式,用摩尔比法确定了PBP与DNA的结合比为2∶1。通过热力学研究得出,在27℃时PBP与鲱鱼精DNA之间相互作用的结合常数为Kθ300.15K=2.16×105L/mol,热力学函数ΔrHθm=-6.18×104J/mol,ΔrSθm=-1.05×103J/(mol·K),ΔrGmθ300.15K=-3.04×104J/mol,结果表明该反应为焓驱动。综合实验结果确定了PBP与DNA的作用方式为部分嵌插以及静电作用。     

一种DNA荧光测定体系的研究与应用

应用钙黄绿素-藏红T为荧光探针,建立了新的DNA测定方法.研究了钙黄绿素-藏红T荧光能量转移体系的荧光性能及其与DNA的结合反应.当pH=8.00时,hsDNA、smDNA和ctDNA的含量与钙黄绿素-藏红T的荧光比值变化量Δ(Fd Fa)具有良好的线性关系,线性范围分别为0.10-5.00、0.50-4.00、0 30-5.00μg mL,检测限(3σ K)分别为0.016、0.021和0.020μg mL;测定DNA合成样品,回收率为91 3%-103.3%.     

A near-infrared fluorescent probe for fluorine ions and its application in the imaging of HepG2 cells

In this study, we have designed and synthesized a new near-infrared (NIR) fluorescent probe (BODIPY-Se) to detect fluorine ions (F-) using a B-Se bond to connect the fluorescent dye BODIPY and benzotrifluoride. The probe exhibited a highly selective fluorescence response to F- with a detection limit of 7.4×108 mol/L. The excitation and emission spectra of the probe in the NIR region avoid background fluorescence interference present in biological systems. The fluorescent imaging of HepG2 cells demonstrated ...     

基于双氧水检测的硼酸酯荧光探针的研究

以邻羟基苯乙酮和苯甲醛为原料经过羟醛缩合反应、氧化反应以及取代反应,合成了一种新型荧光探针,其结构经过1H NMR、MS进行了表征。利用荧光光谱,探讨了该探针对双氧水响应的光谱性能、选择性、p H值以及反应时间等因素。结果表明,该探针能很好地对双氧水进行检测。     

水溶性高分子金属卟啉络合物的合成及其与DNA作用的研究

利用荧光探针技术和圆二色谱等分析手段对两种水溶性高分子金属卟啉络合物同DNA的作用进行了初步研究．实验表明,所合成的高分子金属卟啉络合物在同DNA作用时存在有嵌入模式,金属卟啉络合物大的平面环疏水结构和亲水性基团的引入促进了这种作用．     

一种用于次氯酸根检测的螺二芴类双光子荧光探针的合成及性能研究

设计并合成了一种用于次氯酸根(ClO^-)检测的螺二芴类双光子荧光探针.加入50μmol·L^-1ClO^-后,可以裸眼观察到探针溶液颜色从黄色变为无色,在波长365 nm的紫外光照射下荧光明显增强(10倍).在物质的量浓度为0～20μmol·L^-1范围内,荧光强度(Y)与ClO-物质的量浓度(X)存在较好的线性相关性,其拟合方程为：Y=7.142 41X+65.675 57.根据3σ/k准则,计算出检测限为2μmol·L^-1,表明探针对ClO^-有很好的灵敏性.该探针对ClO^-的荧光响应不受其他干扰物存在的影响,对ClO^-有很好的选择性.探针的荧光响应在广泛的pH范围内表现稳定,适宜在pH值为6～9对ClO^-的检测.用Z-扫描技术,测得探针的双光子吸收截面为300 GM.实验结果表明：该探针对环境和生物中ClO^-的检测具有很好的应用前景.     

A facile and general approach for the synthesis of fluorescent silica nanoparticles doped with inert dyes

A general and facile approach was developed for the synthesis of almost monodisperse fluorescent silica nanoparticles (NPs) doped with inert dyes, which are organic fluorophores that are strongly fluorescent but are hydrophobic or lack a covalent binding group. The prepared NPs were mesoporous and the dye molecules were encapsulated in the pores via hydrophobic interaction with the CTAB template. The NPs were stable and highly fluorescent in aqueous solution, and have potential applications in bioanalysis a...     

Studying the interaction between gyrase and DNA using magnetic tweezers

The DNA gyrase of Escherichia coli plays an essential role in the life of this microorganism.It is unique among all topoisomerases because of its ability to introduce negative supercoils into DNA.This study investigated the single molecular interaction of E.coli gyrase with DNA using magnetic tweezers.The results showed that,in the absence of ATP,gyrase weakly binds the G and T segments.The stretched force of 0.7 pN can gradually destroy the binding,whereas that of 5.9 pN directly destroys it.Addition of high concentrations of norfloxacin enhances gyrase binding to both segments,making them adapt to 5.9 pN.DNA gyrase reduces the plectonemic dimension,which was determined by the bacterial enzyme and not by the pull force.Moreover,it has different affinities for positive supercoils,which it prefers,and negative supercoils.The time distribution of the dissociation of gyrase from DNA has a double-exponential form.We herein propose a model to explain this distribution and compare the results with those of other models.     

光谱法研究诺氟沙星与DNA的作用

利用荧光光谱和紫外光谱研究了诺氟沙星(NFA)与DNA作用.实验结果表明,DNA以静态猝灭的方式使得诺氟沙星的荧光强度显著降低,NFA在浓度为5.86×10-7～1.95×10-6mol·L-1时,体系的荧光强度与DNA的浓度在一定范围内存在着线性关系,其中在NFA浓度为5.86×10-6mol·L-1时,存在最宽的线性范围,DNA的浓度为2.0×10-5～2.22×10-4mol·L-1.确定了最佳反应条件,该方法简便快速,背景干扰小,对于合成样品中DNA测定结果令人满意.     

Green fluorescent protein （GFP） as a vital marker for studying the interaction of Phytophthora sojae and soybean

Transgenic Phytophthora sojae strains that produce green fluorescent protein （GFP） were obtained after stable DNA integration using the Hsp70 promoter and the Ham34 terminator of Bremia lactucae. The expression of GFP during different developmental stages of P. sojae was observed using fluorescent microscopy. Based on this reporter system, the histopathologic events caused by the pathogen in soybean leaves, hypocotyls and roots were monitored. Meanwhile, the difference in resistance between different soybean cultivars against P. sojae was analyzed microscopically in roots. The results indicate that GFP can be stably expressed in zoosporangia, zoospores, cysts, hyphae and oospores of P. sojae. Using the GFP marker, the infecting pathogens in leaves, hypocotyls and roots of host could be distinctly visualized. The germ tube length of cysts germinating on the roots of resistant cultivar Nannong 8848 was longer than that on the roots of susceptible cultivar Hefeng 35. These results show for the first time that this eukaryotic reporter can be used in P. sojae as a stable and vital marker, allowing the study of genetics of this hemibiotrophic pathogen.     

壳聚糖与DNA相互作用的单分子方法研究

利用原子力显微镜与磁镊技术研究不同浓度的壳聚糖与多种DNA分子的作用,对其凝聚形态及力谱曲线进行观察。在恒力情况下,壳聚糖与DNA的电荷比k分别为3和4时,从DNA的力谱曲线上可以看见0.2~0.5μm左右的跳跃阶梯及线性增长区域,原因可能是壳聚糖-DNA聚合物中出现了环状、棒状及球状结构;利用原子力显微镜观察壳聚糖与λ-DNA及bpr322-DNA作用后的凝聚形态,发现随着壳聚糖与DNA电荷比k的增加,球状与棒状聚合物的比例明显减小,而且凝聚物的尺寸也整体减小。最后根据实验结果,建立了壳聚糖导致DNA构象变化的直观模型。     

多巴胺与鲱鱼精DNA相互作用的电化学研究

通过循环伏安法制备了聚L-半胱氨酸修饰电极,利用该修饰电极研究了Cu2+存在下多巴胺（DA）与鲱鱼精DNA在Tris-马来酸缓冲溶液中的相互作用.结果表明:在pH7.0 的Tris-马来酸缓冲溶液中,DA在聚L-半胱氨酸复合膜修饰电极上有一对明显的氧化还原峰,加DNA后氧化还原峰电流明显下降,说明DNA和DA能发生相互作用.再加入Cu2+后氧化还原峰电流减小更大,说明Cu2+对DA和DNA的相互作用有较大影响.     

Enhancement of the immobilization and discrimination of DNA probe on a biosensor using gold nanoparticles

Nanoparticles have been widely utilized in biological research in recent years. 1,6-Hexanedithiol was used as the medium in this work to attach Au nanoparticles on the gold plates. We studied the immobilization and hybridization of DNA probes onto the resulted plates using the quartz crystal microbalance method. Approximately 3–5 times adsorption enhancement of HS-DNA on the plates was obtained under our experimental conditions. Using this DNA sensor, the sensitivity was promoted by approximately three times to 0.35 ώg/mL.     

菠菜基因组DNA提取方法与条件的研究

分别采用CTAB、SDS、高盐低pH三种方法从菠菜叶片中提取基因组DNA,通过A260/A280值、A260/A230值和琼脂糖凝胶电泳对所提取的DNA进行定量、定性分析和综合比较。研究提取缓冲液用量、水浴时间、蛋白质沉淀剂(氯仿-异戊醇)用量、DNA沉淀剂(异丙醇)用量等条件对DNA提取效率和纯度的影响。并对CTAB法的提取条件进行了正交试验,优选出菠菜基因组DNA提取的适宜条件。     

荧光素-番红花红T能量转移体系的研究及其作为核酸荧光探针的应用

研究了以能量转移作为核酸荧光探针的可能性，提出利用能量转移体系测定ＤＮＡ的新方法．该方法灵敏、快速，重现性好，ＤＮＡ测定的线性范围是０．０５～１０ｍｇ／Ｌ，检测限为０．０４７ｍｇ／ＬＣＴＤＮＡ（ｎ＝１１）．     

荧光法研究苯甲酸氮芥铕配合物与DNA的相互作用

 在Tris-HCl(pH7.2)介质中,研究了1个新的苯甲酸氮芥铕(Ⅲ)配合物(Eu(BANM)₃·H₂O)与DNA相互作用的荧光性能,表明此配合物可作为DNA定量测定的荧光试剂.此外以溴化乙锭(EB)为荧光探针,对配合物与DNA的结合机理进行了探讨.结果证明DNA与配合物之间存在插入和沟结合2种非共价作用方式,配合物与DNA的平均结合常数为7.19×10⁵.     

锌(Ⅱ)配合物与DNA作用的研究

采用紫外可见光谱、荧光光谱等光谱方法研究了配离子[zn(phen)2]^2 (phen=邻菲咯啉)与小牛胸腺DNA的相互作用。在DNA存在下，配合物的紫外吸收光谱产生了明显的减色效应。DNA的碱变性曲线在配合物的存在下向pH值增大的方向移动，增色效应减小．荧光光谱表明EB(溴化乙锭)-DNA体系的荧光强度随[zn(phen)2]^2 的加入而迅速减弱，表明锌的邻菲咯啉配合物与DNA之间发生了插入作用。