A novel real - time fluorescent rt - pcr was established to detect pnrsv this technique uses a taqman probe to monitor in real time amplification of target genes , a taqman probe labeled by a quenching and fluorecenting was added to the pcr reaction buffer 另外还设计了taqman荧光探针，确定了检测条件和参数，建立了实时荧光rtwcr方法，该方法是指在pcr反应体系中加入带有荧光基团互补探针，如果有pcr反应（扩增） ，荧光信号就较大。
So the method was accurate and reliable . the in situ nile blue ( nb ) dimer with weak fluorescence in the solution of anionic surfactant sodium dodecylbenzene sulfonate - 6 ( dbss ) was also used as the fluorescence probe for the determination of total proteins in human serum 以阳离子染料耐尔蓝在阴离子表面活性剂十二烷基苯磺酸钠存在下形成的现场弱荧光二聚体作为测定人血清总蛋白的荧光探针。该法用于实际样品的测定，与临床方法测定的结果相吻合。
In view of the above - discussed technological backdrop , it is apparent that the field has urgently awaited the innovative development of long - wavelength and ratiometric fluorescent ca2 + indicators which are concomitantly able to target precisely to specific intracellular locations . in this thesis , the investigations include the following several aspects : 1 新型细胞c扩+荧光探针的合成及性能研究1 .新型钙荧光探针的合成设计合成了两种新型钙荧光探针一stdhi和stdbt (其中stdin已制成相应am酉旨型可导入细胞) 。
On the basis of the mentioned above and the real - time pcr principal , we have designed and optimized the primers and probes , set up the real - time qualitative pcr detection system . meanwhile , using the endogenous zein - maize and lectin - soybean as interior collate gene , epsps and cryia ( b ) as target genes , set up the real - time quantitative pcr detection system 在此基础上，根据荧光pcr原理，设计、优化引物和荧光探针，建立了荧光pcr定性检测方法体系，并以玉米内源基因( zein ) 、大豆内源基因( lectin )作为内标基因，以epsps和cryia ( b )作为目的基因，建立了荧光pcr定量检测方法体系。
. from the direct mutant of spirulina platensis ( sp - d ) , we got high purity and activity phycobiliprotein which could grow crystals . the algae fluorescent probe prepared by coupling the above polyclonal antibody to phycobilipotein not only keeps the property of stronger anti - fluorescence quenching but also has the lower fluorescent background when it was used for labeling stoma cells of pea tendril 以原核表达的peac1为抗原制备了免疫活性较好的抗豌豆肌动蛋白的多克隆抗体，从螺旋藻中纯化了高纯度、高活性、能结晶的藻胆蛋白，将两者偶联制备的藻荧光探针，不仅保持了藻胆蛋白很强的抗荧光淬灭能力，而且用于豌豆卷须气孔细胞荧光标记时有更低的荧光背景。
2 . in order to deeply probe into the influence of crosslinked degree on er properties and expediently characterize the structure of the supramolecular complex , six hydrosoluble - cyclodextrin polymer / 3 - hydroxy - 2 - naphthoic acid er particles were synthesized . fluorescence analysis conformed that all the polymers can form 1 : 1 type supramolecular complex with the guest 为了进一步考察交联度对电流变效应的影响，并便于超分子配合物颗粒的结构表征，制备了6种不同交联度的水溶性-环糊精聚合物，以3 -羟基- 2 -萘甲酸为荧光探针探讨了不同交联度超分子配合物的组成、稳定性及荧光性质。
Although fluorescence labeling method was widely used in the observation of microfilaments " dynamic distributions in vivo , all of the traditional methods have their limits . the use of gfp fusion and the preparation of new type of algae fluorescent probe will facilitate our further study on plant actins 荧光标记技术虽然广泛用于细胞内微丝骨架分布及其动态的研究，但传统的荧光标记方法用于植物肌动蛋白研究时都具有一定的局限性，利用gfp融合以及研制新型的藻荧光探针有助于对植物肌动蛋白的深入了解。
The indicators quin - 2 , indo - land fura - 2 have the advantage of wavelength - shift upon binding ca2 + but share a common drawback in that the excitation bands for these indicators occur at uv wavelength ( 340 - 360nm ) , where autofluorescence from cells is high , radiation - damage of cellular structures is serious , the transmission of microscope optics is poor , and the availability of aberration - corrected optics for confocal microscopy is limited 迄今为止，尚未见既可长波激发、又与ca ~ ( 2 + )结合后荧光峰位移、且对胞内ca ~ ( 2 + )测定具特异性、可在亚细胞水平进行ca ~ ( 2 + )测定的整体性能优良的钙荧光探针及其在生命科学中的应用研究报道。
The loop sequence of mb1 and mb2 were the anti sense and sense sequence ofing1 , respectively the sequence of mb3 was a piece of ssrna sequence in tobacco mosaic virus , which had no analogical to human gene . mbl was the most suitable probe because mbl had the highest fluorescence enhancemen after hybridizing wtth rna extrated froin normal cell 第三章，根据一种常见的病毒烟草花叶病毒( tmv )的核酸序列设计了分子信标荧光探针，由于tmv的遗传物质是rna ，分子信标又具有很高的特异性和灵敏度，因此感染了病毒粒子的植物叶片在经过简单处理后，可用分子信标检测叶片上。
Not only can it be excited by visible light but also displays a spectrum - shift upon binding with ca + . moreover , it targets precisely into cytosol only , and thereby exhibits great advantage beyond flou - 3 and ruro - 2 . thus it is possible to use stdin - am to measure real - time variation of [ ca2 + ] in cytosol 同时细胞荧光图像分析显示， stdhi一am进入细胞后只标记胞浆c扩+而不标记胞核c扩+ ，是目前唯一的可以am酉旨型无损伤导入细胞、双波长、可见光激发的胞浆特异性c扩+荧光探针。
