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荧光素

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In this dissertation, firstly, the conditions of isolation and purification of specific toxin fractions produced by Exserohilum turcicum has been studied, and a high-efficient method has been established;secondly, the toxins were analysed by UV-Vis spectrophotometry;then the effect of specific toxin on death rate of corn leaves protoplast was studied by FDA as a tinct reagent;finally, using ANS as a probe and SDS-PAGE to study the membrane protein of the corn leaves with Htl gene. In a word, the aim of the research is to search some existent evidences of the toxin binding site on protoplasmic membrane, and provide proofs for nosogenesis of specific toxin in terms of cytology and the molecular biology.

本研究对玉米大斑病菌1号小种毒素的特异性组分进行分离,建立了高效的特异性毒素分离纯化方法,并对其进行了紫外波谱分析;用荧光素双醋酸酯染色法,测定了毒素特异性组分对原生质体死亡率的影响;进而用1-苯胺基-8-萘磺酸荧光探针标记法和SDS-PAGE电泳法对特异性毒素组分与Htl基因玉米细胞互作过程中的质膜蛋白进行了研究,旨在证明Htl基因玉米细胞质膜上存在特异性毒素组分结合蛋白,为从细胞和分子生物学角度阐明毒素的致病机理提供证据。

METHODS: The BEC were preincubated with crocetin (1, 0.1 and 0.01 μmol/L) for 12 h, then exposed to AGE (100 mg/L). The RAGE mRNA expression was detected by RT-PCR analysis, the intercellular adhesion molecule-1(ICAM-1) was measured by ELISA. The extracellular superoxide ion and thiobarbituric acid reactive substances were assessed with superoxide ion kit and colorimetric assay, respectively The intracellular I2O2 was also detected using the probe 2,7-dichlorofluorescein. The mitochondrial membrane potential and mitochondrial Succinate dehydrogenase were analyzed by the retention of rhodamine 123 (Rh123) and MTT.

不同浓度的西红花酸(1、0.1、0.01μmol/L)预孵BEC细胞12h后,用AGE(100mg/L)刺激细胞12h,RT-PCR法测定RAGEmRNA的表达水平;ELISA法测定细胞间黏附分子-(ICAM-1)的表达;试剂盒分别检测胞外超氧阴离子和硫代巴比妥酸反应产物浓度;同时,还用2,7-二氯荧光素测定了胞内H2O2的浓度,并用罗丹明123(Rh123)荧光法及MTT法分别检测细胞线粒体膜电位水平和其琥珀酸脱氢酶的活性。

Fluorescein diacetate, which generates fluorescence upon hydrolysis, is used as the assay substrate.

荧光素双乙酸盐在水解时产生荧光,可用于培养基化验。

Fluorescin sodium ; fluorescence intensity ; intensity of illumination ; temperature ; mass concentration

荧光素钠;荧光强度;光照强度;温度;质量浓度

Objective To investigate the characteristics of fundus photography and fundus fluorescein angiography of IRVAN (idiopathic retinal vasculitis, aneurysms, and neuroretinitis) syndrome and Eales disease.

目的 观察特发性视网膜血管炎、动脉瘤、视神经视网膜综合征和Eales病荧光素眼底荧光血管造影特征的异同。

The firefly and Renilla Dual-Glo Luciferase Assay System was purchased from Promega.

荧光剂和Renilla 二元球蛋白(TM荧光素酶检测系统从普洛麦公司购买

First of all, the cytokeratin 19 promoter segment was cloned from hepatocellular carcinoma cell line HepG2 and then following the promoter a renilla luciferase and a red fluorescence protein's fusion gene were inserted to finish the double report lentiviral vector.

首先,通过PCR方法从肝癌细胞系HepG2的全基因组中克隆了CK19启动子片段,构建了CK19启动子调控的海肾荧光素酶和红色荧光蛋白(red fluorescent protein,RFP)双报告载体(pSicoR-CK19-hrl-mrfp)。

Cells were immediately fixed and processed for immuno?uorescence and tested for translocation ofβ-catenin.β-catenin and target genes of Wnt/β-catenin signaling COX-2, cyclinD1, c-fos, c-Jun mRNA expression were examined by RT-PCR. Saos-2 osteoblastic cells were co-transfected with a wild type or mutant Tcf luciferase reporter gene and Renilla luciferase plasmid. Posttransfection cells were subjected to 1 hour of 12% elongation.

作用1h、2h和4h后,用免疫荧光的方法检测β-catenin的表达和在细胞上定位的变化;用半定量RT-PCR检测β-catenin mRNA表达的变化;用质粒转染的方法检测荧光素酶报告基因Tcf转录的情况;用半定量RT-PCR检测Wnt/β-catenin通路下游基因COX-2,cyclinD1, c-fos, c-Jun mRNA表达的情况;用免疫共沉淀的方法检测β-catenin和E-Cadherin的结合情况。

To improve the stability of small unilamellar vesicles in vitro, the permeability of SUV from soybean phosphatidylcholine coated by collagen and chitosan was studied respectively by using 5(6)-Carboxyflourescein5(6-CF as a fluorescence probe.

为了提高小单层脂质体(SmalUnilamelarVesicles,SUV)的体外稳定性,以5(6)-羧基荧光素(5(6)-CF)为荧光探针,研究了脂质体被胶原蛋白和壳聚糖这两种天然生物大分子包覆后的通透性。

They have been used in the detection of a variety of protein, nucleic acid, cell, HIV-1 nucleocapsid etc.

随着生物技术的不断发展,基于荧光素的荧光探针已经在蛋白质、核酸、细胞检测及免疫分析等方面发挥了重要作用。

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