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1.
A novel, homogeneous and sensitive assay for the detection of single nucleotide polymorphisms (SNPs) by integration of rolling circle amplification (RCA) and cationic conjugated polymer (CCP) has been developed and tested. Mutant DNA serves as the template for specifically circularizing a padlock probe (PLP) with a sequence that is complementary to the mutant DNA. Afterwards, the mutant DNA directly acts as the primer to initiate the RCA reaction in the presence of phi29 DNA polymerase that generates a long, tandem single-strand DNA product. During the RCA reaction, fluorescein-labeled dUTPs are incorporated into the RCA products. When the CCP is introduced, efficient FRET from CCP to fluorescein occurs as a result of the strong electrostatic interactions between the CCP and the DNA produced by RCA. The wild-type DNA contains a single base mismatch with PLP with the result that the PLP is not circularized, RCA is not triggered and inefficient FRET results. By measuring the change of the emission intensities of CCP and fluorescein, it was possible to detect the SNP in a homogeneous manner. The method is sensitive and specific enough to detect 0.1 pmol/L mutant DNA and to determine a mutant allele frequency as low as 2.0%.  相似文献   

2.
基于水溶性氟硼二吡咯类共轭聚合物PBF可与长波长区(> 600 nm)荧光染料产生荧光共振能量转移(FRET),结合引物延伸反应,发展了一种均相检测miRNA的新方法.方法首先通过引物延伸反应在目标miRNA分子上引入荧光染料Cy5,形成miRNA-Cy5. 然后,PBF与miRNA-Cy5/DNA杂合体通过静电力结合并发生从PBF到Cy5的有效FRET,实现了基于阳离子共轭聚合物(CCP)的长波长区miRNA的均相检测,方法灵敏度高、特异性好,测定miR-221的线性为15~6 000 pmol/L,检出限(3σ)为8.4 pmol/L.方法拓展了CCP的应用,为基于CCP的生物传感和生化分析提供了新的均相检测平台.  相似文献   

3.
The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energy transfer (FRET) assay for multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers (CCP). The newly established detection system could detect as low as 2% mutant DNAs in DNA admixtures. By triggering the emission intensity change of CCP and the dyes labeled in the DNA, four possible statuses (three mutations and one wildtype) can be differentiated in one extension reaction. The detection efficiency of this new method in clinical molecular diagnosis was validated by determining KRAS and BRAF mutations of 51 formalin-fixed paraffin-embedded (FFPE) ovary tissue samples. Furthermore, the result of the CCP-based multi-step FRET assay can be directly visualized under UV light so that no expensive instruments and technical expertise are needed. Thus, the assay provides a sensitive, reliable, cost-effective and simple method for the detection of disease-related gene mutations.  相似文献   

4.
聚合物驱后期阳离子聚合物堵聚可行性研究   总被引:2,自引:0,他引:2  
室内实验结果表明,阳离子聚合物与阴离子聚合物(部分水解聚丙烯酰胺)发生反应可产生稳定的絮状沉淀,完全反应时阳、阴离子聚合物的质量比为30:1;当絮状沉淀的浓度较高时,它具有很好的携砂能力,絮状沉淀可使岩心的渗透率降低88%以上,且具有良好的抗冲刷能力。矿场试验表明,采用此种阳离子聚合物对产出液中阴离子聚合物浓度高的油井进行封堵,具有明显的增油、降水效果。  相似文献   

5.
室内实验结果表明 ,阳离子聚合物与阴离子聚合物 (部分水解聚丙烯酰胺 )发生反应可产生稳定的絮状沉淀 ,完全反应时阳、阴离子聚合物的质量比为 30∶1;当絮状沉淀的浓度较高时 ,它具有很好的携砂能力 ,絮状沉淀可使岩心的渗透率降低 88%以上 ,且具有良好的抗冲刷能力。矿场试验表明 ,采用此种阳离子聚合物对产出液中阴离子聚合物浓度高的油井进行封堵 ,具有明显的增油、降水效果。  相似文献   

6.
以红薯淀粉为原料进行化学改性,合成了一种阳离子型高分子絮凝剂F4,并考察了各因素对合成的影响.研究结果表明,接枝共聚的最佳合成条件为:淀粉5g,引发剂0.35g,丙烯酰胺10g,反应温度50℃,反应时间3h.阳离子化条件为:氢氧化钠0.08g,阳离子醚化剂3g,反应温度70℃,反应时间2h.  相似文献   

7.
共轭多孔聚合物(CPPs)具有扩展的共轭体系和多孔性,是一种具有广阔应用前景的非均相无金属光催化剂.本文中,我们报道一种基于噻吩的有机多孔聚合物的设计和制备,首先利用Suzuki偶联反应合成出单体3,3',5,5'-四(噻吩-2-基)-1,1'-联苯,再以三氯化铁为催化剂催化噻吩偶联,成功制备一种共轭有机多孔聚合物(PTB).这种聚合物光吸收可达到800 nm,比表面积为699.8 m~2·g~(-1),并且中孔和微孔共存.在非均相体系中,这种催化剂对胺氧化偶联表现出有效的光催化性能,反应产率可达94%,并且亚胺为其唯一产物.  相似文献   

8.
天然改性阳离子絮凝剂制备及性能研究   总被引:4,自引:0,他引:4  
介绍了以玉米芯为母体,以阳离子醚化剂为改性剂,合成阳离子型改性讷发子絮凝剂CNPF的方法,并对该絮凝剂的性能进行了研究,通过正交实验选择最佳条件,反应温度50℃、时间3h。配料比:0.03mol阳离子试剂/5g玉米芯粉。同时以高岭土悬浊液为絮凝效果考察体系,探讨了CNPF的絮凝性能。结果表明,CNPF絮凝效果优于6水氯化铝,在实际应用中,对粘土污水体系的处理表明,CNPF具有良好的抗干扰能力。  相似文献   

9.
分析了回波信号以及其共轭信号的FRFT幅值特性,并利用此特性提出了一种基于共轭FRFT模函数对消的海面运动目标检测方法。该方法利用回波信号在最佳变换角度有较好的能量聚集,而其共轭信号在此最佳变换角度没有能量聚集。利用两者FRFT模函数之差对消部分杂波,在保证信号能量基本不被削弱的前提下达到抑制杂波、提高检测概率的目的。高斯噪声与IPIX雷达数据实验表明该方法在低信杂比条件下有较好的检测性能。  相似文献   

10.
A novel light-emitting alternating copolymer, known as hyperbranched conjugated polymer (HPV), has been designed and synthesized by Wittig reaction, and so have its linear counterparts, including MPV and PPV. FTIR, 1H NMR, 13C NMR and GPC were combined to characterize these polymers. Their photophysical properties were investigated both in solution and in solid film. Interestingly, almost the same maxima of absorption and emission wavelengths were observed for the solution of HPV and MPV in CH2Cl2 respectively, located at the shorter wavelength region related to PPV. In addition, the absorption peak of HPV film blue shifted while its emission peak red shifted as MPV compared with the case of its CH2Cl2 solution. However, both the absorption and emission spectra of the PPV film showed red shift, which resulted from a special electronic structure.  相似文献   

11.
以对苯二乙腈和对羟基苯甲醛为原料通过Knoevenagel缩合反应合成α,α’-双-(对羟基亚苄基)-对苯二乙腈(M1),然后与环氧氯丙烷通过Williamson反应合成α,α’-双-[对-(2,3-环氧丙氧基)亚苄基]-对苯二乙腈(M2). 以2-氰基-2-丙基苯并二硫为可逆加成断裂链转移试剂,引发2-(2',3',4',6'-四-O-乙酰基-α-D-吡喃甘露糖基氧基)甲基丙烯酸乙酯聚合制备了含双硫酯片段的聚合物P1,P1与M2通过“一锅法”合成含对苯二乙腈单元聚合物P2,脱除乙酰基保护得到水溶性荧光聚合物P3. 利用核磁共振波谱、傅里叶变换红外光谱、凝胶渗透色谱、紫外-可见光谱和荧光光谱对化合物和聚合物的化学结构和光学性质等进行了表征与测试. 结果表明:P3的紫外吸收峰和荧光发射峰分别在368 nm和440 nm,P3荧光强度随着DMSO-H2O体系DMSO体积分数的增加而逐渐增强,并且在体积分数大于80%时荧光强度显著增强,表现出聚集诱导发光特性. P3具有较好的热稳定性,相转变温度为118 ℃.  相似文献   

12.
阳离子聚合物处理油田采油污水研究   总被引:7,自引:2,他引:7  
为了使油田含油污水处理后能达到回用标准,通过阳离子聚合物与聚合铝复配对油田采油污水处理进行了研究.结果表明聚合物分子量为120万,加入浓度为0.5~1.0mg/L时,与聚合铝复配后,对采油污水中的悬浮物、含油量及CODcr的去除率分别达到99.7%,97.7%及92.0%,并使处理后水中悬浮物及含油量分别降低到1.2mg/L及5.0mg/L,达到了油田回用水水质标准.复配体系对含油污水的处理效果受污水pH值及温度的影响,但影响比单独使用聚合铝时小.  相似文献   

13.
Fullerenes and functionalized fullerenes have at-tracted extensive attention of chemists, material scientists and biologists since the discovery of [60]fullerene (C60). Functionalized fullerenes exhibit a wide range of biologi-cal and medicinal activities, such as DNA photocleavage, antioxidants, neuroprotective agents, antiapoptotic activity, enzyme inhibition, anti-HIV activity, contrast agents and radiotracers[1—3] owing to their unique photochemical, radical-quenching and hydrophobic pr…  相似文献   

14.
为了得到一种新型高效的聚合物太阳能电池材料,通过Stille聚合反应合成了一种以噻唑并噻唑为电子受体单元和硅基联噻吩为电子给体单元的交替共轭聚合物(PTTz-Si)。这种聚合物具有较窄的光学带隙(1.77 eV)、较高的热稳定性以及比较宽泛的紫外可见吸收光谱,其良好的溶解性保证了可以通过溶液加工制备成有机太阳能电池器件,是一种潜在的聚合物太阳能电池活性层供体材料。通过核磁共振氢谱、碳谱、热重分析、紫外可见吸收光谱、凝胶渗透色谱和电化学等测试方法对该聚合物进行了表征,并且将聚合物与PC71 CM共混制备聚合物太阳能电池器件,获取0.76%的光电转化效率。  相似文献   

15.
Redesign of the coenzyme specificity of a dehydrogenase by protein engineering   总被引:61,自引:0,他引:61  
N S Scrutton  A Berry  R N Perham 《Nature》1990,343(6253):38-43
Directed mutagenesis and molecular modelling have been used to identify a set of amino-acid side chains in glutathione reductase that confer specificity for the coenzyme NADP+. Systematic replacement of these amino acids, all of which occur in a 'fingerprint' structural motif in the NADP+-binding domain, leaves the substrate specificity unchanged but converts the enzyme into one displaying a marked preference for the coenzyme NAD+.  相似文献   

16.
阳离子聚合物/膨润土对苯酚的吸附及其机理研究   总被引:4,自引:1,他引:4  
采用阳离子聚合物聚环氧氯丙烷二甲铵(EPI-DMA)和聚二甲基二烯丙基氯化铵(PDMDAAC)分别对钠基膨润土进行了改性,研究了改性膨润土吸附苯酚的机理及其主要影响因素。实验结果表明,膨润土经阳离子聚合物改性后,吸附苯酚的能力大大提高。振荡时间、温度、pH值、离子浓度对阳离子聚合物/膨润土吸附苯酚的性能均有一定的影响。PDMDAAC/膨润土和EPI-DMA/膨润土分别在振荡时间为20min和60min时,达到吸附平衡;温度升高,吸附量下降,不利于吸附反应的进行;pH相似文献   

17.
18.
中原油田是一个多层系、多油藏类型的复杂断块油田 ,部分区块储层中黏土特别是膨胀性黏土含量高 ,对注入水水质的要求较高 .本文对阳离子聚合物与聚合铝复配处理中原油田采油污水进行了研究 ,结果表明 :当聚合铝浓度为 5 0 mg/L时 ,阳离子聚合物的分子量、加入浓度、污水的 p H值、污水温度等对处理效果影响大 ,当阳离子聚合物的黏均分子量为 1 2 0万、加入浓度 3~ 5 mg/L、污水的 p H值为 6.5~ 8.5、污水温度在 45℃时 ,中原油田采油一厂采油污水中悬浮物、油的含量分别由处理前的 1 65 mg/L,1 1 8mg/L降低到 1 .2 mg/L及 5 .0 mg/L ,滤膜系数达到 35  相似文献   

19.
阴、阳离子聚合物交替注入用于注聚区封窜和调剖   总被引:3,自引:0,他引:3  
胜利孤岛油田注聚区块过早见聚,为封堵聚合物从油井窜流产出和调整注聚剖面,研制出阳离子调剖剂NCP。室内试验结果表明,向用1000mg/L的HPAM驱油后的岩心中注入1.5×10~4~3.0×104mg/L的NCP1.0PV,在70℃恒温24h后的堵油率大于96%,突破压力梯度2.6~3.3MPa/m。注入2.0×10~4mg/L NCP溶液的岩心,连续注入50PV含800mg/LHPAM的地层水,水相渗透率持续下降;在岩心试验中,堵水率随NCP浓度的增大而增大。结果表明:NCP可用于注聚区块调整注聚剖面,在孤岛注聚区进行的5井次封窜试验中,3口井收到良好效果。  相似文献   

20.
单态氧是一种重要的高活性氧自由基,在材料科学,生物以及医学等领域具有重要的研究价值.本文阐述了近年新兴的商品化单态氧荧光探针(Singlet Oxygen Sensor Green reagent,SOSG)的光谱特性和检测原理,并综述了其在单态氧检测应用情况.最后,展望了SOSG在单态氧检测应用研究的发展趋势.  相似文献   

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