title: Inhibition of G-quadruplex assembling by DNA ligation: a versatile and non-covalent labeling strategy for bioanalysis.
abstract: Through tuning relative thermodynamic stabilities (I, II and III), DNA ligation was coupled to split G-quadruplex probes and a versatile, non-covalent labelling and fluorescent strategy was constructed based on inhibition of template-directed G-quadruplex assembling by ligation reaction. The non-covalent complex between G-quadruplex and fluorescent probe was employed as signalling label and thus covalent modification of DNA probes with fluorescent probes was avoided. Selective detection of small biomolecules (ATP and NAD(+)) in the nanomolar range was realized due to the cofactor-dependent activity of DNA ligases (T4 and Escherichia coli DNA ligase). By virtue of the simple strategy, the effect of mismatch position of single-base mismatched template DNA on the ligation efficiency was validated. Meanwhile, highly mismatch-influenced ligation efficiency of ligase endows the cost-effective strategy great potential for single-nucleotide polymorphism (SNP) analysis. The non-covalent labeling strategy provides a versatile and cost-effective platform for monitor of DNA ligation, cofactor detection, SNP analysis and other ligase-based assays.
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Jiangtao
authors.initials: J
authors.lastname: Ren
authors.forename: Jiahai
authors.initials: J
authors.lastname: Wang
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Erkang
authors.initials: E
authors.lastname: Wang
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis*; D017122:Bacteriophage T4 / Q000201:enzymology; D015374:Biosensing Techniques / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D054856:G-Quadruplexes; D009243:NAD / Q000032:analysis*; D020641:Polymorphism, Single Nucleotide; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D009243:NAD; D000255:Adenosine Triphosphate; D004247:DNA; D011088:DNA Ligases
keywords: DNA ligation; Fluorescent probe; G-quadruplex; Non-covalent labeling