Luminescent detection of nicking endonuclease Nb.BsmI activity by using a G-quadruplex-selective iridium(III) complex in aqueous solution

被引:13
作者
Dong, Zhen-Zhen [1 ]
Lu, Lihua [1 ,2 ]
Wang, Wanhe [1 ]
Li, Guodong [3 ]
Kang, Tian-Shu [3 ]
Han, Quanbin [4 ]
Leung, Chung-Hang [3 ]
Ma, Dik-Lung [1 ,5 ]
机构
[1] Hong Kong Baptist Univ, Dept Chem, Kowloon Tong, Hong Kong, Peoples R China
[2] Qingdao Agr Univ, Coll Chem & Pharmaceut Sci, Qingdao 266109, Peoples R China
[3] Univ Macau, Inst Chinese Med Sci, State Key Lab Qual Res Chinese Med, Macau, Peoples R China
[4] Hong Kong Baptist Univ, Sch Chinese Med, Kowloon Tong, Hong Kong, Peoples R China
[5] Hong Kong Baptist Unvers, Inst Res & Continuing Educ, Shenzhen 518057, Peoples R China
来源
SENSORS AND ACTUATORS B-CHEMICAL | 2017年 / 246卷
基金
中国国家自然科学基金;
关键词
Iridium(III) complex; Nicking endonuclease; Luminescence; G-quadruplex; Probe; CYCLOMETALATED IR(III) COMPLEXES; LABEL-FREE DETECTION; SWITCH-ON DETECTION; SIGNAL AMPLIFICATION; STRAND DISPLACEMENT; DNA DETECTION; FLUORESCENCE; ASSAY; STRATEGY; LIGANDS;
D O I
10.1016/j.snb.2017.02.156
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A cyclometalated iridium(III) complex has been synthesized as a novel luminescent probe for G-quadruplex (G4) DNA in aqueous solution, and a G4-based assay was employed for the detection of nicking endonuclease activity using Nb.Bsml as a model enzyme. In the assay, an initial hairpin DNA (5'-TG(4)AG(3)TG(4)AG(3)TG(4)A(2)G(3)A(2)TGCTA(3)TAGCAT(2)C(3)T(2)C(4)AC-3') sequence containing the Nb.Bsml recognition site in its stem region and a partially caged G-rich DNA sequence at its 5'-terminus could be cleaved by Nb.BsmI, which liberates the G-rich DNA sequence and enhances the luminescence of complex 1. This assay showed a linear relation between luminescent signal and Nb.Bsml concentration over the range of 0-10 U/mL (R-2=0.9937) with a detection limit for Nb.Bsml of 0.042 U/mL. Furthermore, this assay showed high selectivity for Nb.Bsmi over other nicking endonucleases, and functioned effectively in the presence of cell lysates. (C) 2017 Elsevier B.V. All rights reserved.
引用
收藏
页码:826 / 832
页数:7
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