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91.
目的:构建重组质粒作为标准阳性模板,建立食品中沙门氏致病菌实时荧光定量聚合酶链式反应检测方法。方法:以致病性沙门氏菌inv A基因上特异性片段为目标,设计并合成引物和Taq Man探针,将目标片段连接到PGM-T载体上构建重组质粒,建立实时荧光定量检测体系,并考察方法的灵敏性、特异性、重复性和准确性。结果:构建出致病性沙门氏菌特异性基因片段的重组质粒,能够作为实时荧光定量聚合酶链式反应检测方法的标准阳性模板,标准曲线方程为Y=-3.151 lg X+42.86(R2=0.999),灵敏度80拷贝反应体系,能特异区分沙门氏菌与类型的细菌,同时,批内和批间的变异系数均小于5%,具有良好的重复性。结论:本方法能够实现对食品中致病性沙门氏菌进行定性定量检测。 相似文献
92.
93.
Internalization and Accumulation in Dendritic Cells of a Small pH‐Activatable Glycomimetic Fluorescent Probe as Revealed by Spectral Detection
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Prof. Zoran Arsov Prof. Urban Švajger Prof. Janez Mravljak Prof. Stane Pajk Anita Kotar Dr. Iztok Urbančič Prof. Janez Štrancar Prof. Marko Anderluh 《Chembiochem : a European journal of chemical biology》2015,16(18):2660-2667
DC‐SIGN, an antigen‐uptake receptor in dendritic cells (DCs), has a clear role in the immune response but, conversely, can also facilitate infection by providing entry of pathogens into DCs. The key action in both processes is internalization into acidic endosomes and lysosomes. Molecular probes that bind to DC‐SIGN could thus provide a useful tool to study internalization and constitute potential antagonists against pathogens. So far, only large molecules have been used to directly observe DC‐SIGN‐mediated internalization into DCs by fluorescence visualization. We designed and synthesized an appropriate small glycomimetic probe. Two particular properties of the probe were exploited: activation in a low‐pH environment and an aggregation‐induced spectral shift. Our results indicate that small glycomimetic molecules could compete with antigen/pathogen for binding not only outside but also inside the DC, thus preventing the harmful action of pathogens that are able to intrude into DCs, for example, HIV‐1. 相似文献
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95.
Yoshiyuki Ohiro Hiroshi Ueda Norio Shibata Teruyuki Nagamune 《Journal of Bioscience and Bioengineering》2010,109(1):15-19
We describe a homogeneous competitive immunoassay for a phosphorylated protein antigen. The assay takes advantage of the enhanced fluorescence resonance energy transfer (FRET) technology, which has a unique characteristic that the FRET signal is increased by the specific interaction of two fluorolabeled leucine zippers. We chose extracellular signal-regulated kinase (ERK) as a model antigen and constructed two molecular probes in which either anti-phosphorylation site antibody or the antigen peptide was chemically conjugated to the enhanced FRET probes. While these molecular probes indicated sufficient FRET signal without antigen, they displayed a significant change in the fluorescent spectrum by mixing with phosphorylated antigens. With this competitive enhanced FRET immunoassay, a phosphorylated ERK concentration within the range from 15 nM to 250 nM could be determined. Because the assay is very simple, it would be applied to not only in vitro assay but also in vivo detection of protein phosphorylation. 相似文献
96.
Dr. Alexander J. M. Disney Prof. Barrie Kellam Dr. Lodewijk V. Dekker 《ChemMedChem》2016,11(9):972-979
The natural product staurosporine is a high‐affinity inhibitor of nearly all mammalian protein kinases. The labelling of staurosporine has proven effective as a means of generating protein kinase research tools. Most tools have been generated by acylation of the 4′‐methylamine of the sugar moiety of staurosporine. Herein we describe the alkylation of this group as a first step to generate a fluorescently labelled staurosporine. Following alkylation, a polyethylene glycol linker was installed, allowing subsequent attachment of fluorescein. We report that this fluorescein–staurosporine conjugate binds to cAMP‐dependent protein kinase in the nanomolar range. Furthermore, its binding can be antagonised with unmodified staurosporine as well as ATP, indicating it targets the ATP binding site in a similar fashion to native staurosporine. This reagent has potential application as a screening tool for protein kinases of interest. 相似文献
97.
芳香多羧酸常用于构筑金属有机框架(MOFs)材料,为了增加MOFs材料的比表面积,从而提高材料对气体的吸附性能,分别以1,3,5-三溴苯和9,10-二溴蒽为原料,通过Suzuki偶联反应设计合成两个芳香多羧酸配体,即3,3'-(9,10-蒽二基)-二苯甲酸和三-(3'-羧基苯基)苯(H3TCPB),其中前者为新化合物,其结构经1HNMR、13CNMR和HRMS表征。由于含蒽材料具有强荧光性能,研究了3,3'-(9,10-蒽二基)-二苯甲酸的固体室温UV/Vis和荧光激发光谱。结果表明,该化合物的最大吸收λmax为252和366 nm,最大发射波长为428 nm,具有发紫光的性质。使用配体H3TCPB和锌盐水热合成一个MOF,即{[Zn4(TCPB)2(OH)2(CH3OH)(DMF)3]·2DMF·H2O}∞。BET比表面积测试结果表明,其比表面积为290 m2/g。 相似文献
98.
合成了1-(邻氯苯基)-3-(2-羟基苯基)-丙二酮(L1)、1-(间氯苯基)-3-(2-羟基苯基)-丙二酮(L2)和1-(对氯苯基)-3-(2-羟基苯基)-丙二酮(L3)3种配体,并将此3种配体分别与 Eu(III)反应生成3种新的稀土配合物。运用元素分析、红外光谱与荧光光谱等手段对配合物进行了表征。结果表明:3种配合物的组成分别为 Eu(L1)3·2H2O、Eu(L2)3·2H2O 和 Eu(L3)3·2H2O。荧光光谱显示,3种配合物的配体均能将吸收的能量有效地传递给三价铕离子,从而使配合物发射出强的铕离子的特征荧光。在3种配合物中,Eu(L1)3·2H2O 的荧光强度远大于 Eu(L2)3·2H2O 和 Eu(L3)3·2H2O 的荧光强度,这说明配体 L1与 Eu(III)离子的能级匹配较好,能量传递效率较高。 相似文献
99.
Jianzhao Liu Jacky W. Y. Lam Ben Zhong Tang 《Journal of Inorganic and Organometallic Polymers and Materials》2009,19(3):249-285
Aggregation generally quenches the light emissions of chromophoric molecules. In this review, we demonstrate that 1,1-disubstituted
2,3,4,5-tetraphenyl siloles and 2,5-difunctionalized siloles as well as their polymers exhibit the opposite behaviors. Instead
of quenching, aggregation has greatly boosted their photoluminescence quantum yields by up to two orders of magnitude, turning
them from faint fluorophores into strong emitters. Such “abnormal” phenomenon of “aggregation-induced emission (AIE)” is attributed
to restricted intramolecular rotations of the peripheral phenyl rings against the central silole core, which block the nonradiative
channel via the rotational energy relaxation processes and effectively populate the radiative decay of the excitons. Utilizing
such a novel effect, siloles and their polymers find an array of applications as: sensors for chemicals, explosives, pH, and
biomacromolecules (proteins, DNAs and RNAs), indicators for determining CMC and monitoring layer-by-layer self-assembling,
biocompatible fluorogens for cell imaging, visualizing agent for DNA gel electrophoresis, biolabels for immunoassay, stimuli-responsive
organic nanomaterials, magnetic fluorescent nanoparticles for potential bio-imaging and -separation, and outstanding materials
for efficient OLEDs and PV cells.
相似文献
Ben Zhong TangEmail: |
100.
概述了荧光指示剂测定细胞内金属离子的原理,仪器及进展。简要介绍了Fura-2/AM,Mag-Fura-2/AM,SBFI和PBFI分别用于细胞内Ca^2^+,Mg^2^+,Na^+和K^+的测定。 相似文献