Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity
文献类型：期刊论文
| 作者 | Zhang, Dapeng; Fu, Rong; Zhao, Qiang; Rong, Haiqin; Wang, Hailin |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2015-05-05 |
| 卷号 | 87期号:9页码:4903-4909 |
| 英文摘要 | Fluorescence anisotropy is a homogeneous, sensitive, ratiometric, and real-time analytical technology. However, it is a great challenge to produce a large fluorescence anisotropy change upon the presence of target small molecules without nanoparticles-dependent amplification. This work reports a nanoparticle-free and multiple G-enhanced fluorescence anisotropy assay for detection of DNAzyme activity. A Pb2+-dependent GR-5 DNAzyme was used as a model. We hybridized the rA-cleavable substrate strand containing a TMR label at the 5'-end with the DNAzyme strand containing an extended three G bases at the 3'-end. By this design, we demonstrate that both fluorescence quenching and the enhanced DNAzyme activity contribute to a Pb2+-induced large fluorescence anisotropy change (vertical bar Delta r vertical bar = 0.168). The limit of detection for Pb2+ is estimated to be about 100 pM with a dynamic range from 200 pM to 100 nM. The interference from the other nine divalent metal ions of 1000-times excess amount is negligible. Moreover, we show an extended assay for evaluation of the interactions of Pb2+ with cysteine and glutathione by the detection of GR5 DNAzyme activity. Collectively, we developed a novel fluorescence anisotropy amplification assay, enabling us to detect DNAzyrne activity and associated cofactors and inhibitors and to characterize the Pb2+-chelation capability of free thiols. |
| 研究领域[WOS] | Chemistry, Analytical |
| WOS记录号 | WOS:000354153600045 |
| 公开日期 | 2016-03-15 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/32628]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Zhang, Dapeng,Fu, Rong,Zhao, Qiang,et al. Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity[J]. ANALYTICAL CHEMISTRY,2015,87(9):4903-4909. |
| APA | Zhang, Dapeng,Fu, Rong,Zhao, Qiang,Rong, Haiqin,&Wang, Hailin.(2015).Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity.ANALYTICAL CHEMISTRY,87(9),4903-4909. |
| MLA | Zhang, Dapeng,et al."Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity".ANALYTICAL CHEMISTRY 87.9(2015):4903-4909. |
入库方式： OAI收割
来源：生态环境研究中心
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