Reference - Detail
RRC ID | 3811 |
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Author | Yoshitake K, Waki S, Ueda H. |
Title | Dimerization-based homogeneous fluorosensor proteins for the detection of specific dsDNA. |
Journal | Biosens Bioelectron |
Abstract |
While there are many hybridization-based DNA sensors, none of them can detect native double-stranded DNA (dsDNA), which is most commonly found in physiological conditions. Here we made novel fluorosensor proteins comprised of a pair of two zinc fingers with an N-terminal dimerization motif and a C-terminal GFP variant to detect specific dsDNA sequence in a homogeneous solution. When a pair of purified zinc finger-GFP color variant proteins (Zif12-eCFP, Zif12-eYFP) were mixed and added with specific dsDNA with 12 bp inverted repeat (IR), fluorescence spectra of the solution showed significant concentration-dependent enhancement of fluorescence resonance energy transfer (FRET), with the detection limit of approximately 10nM. No significant change in FRET was observed if nonspecific DNA was added, indicating dsDNA-dependent dimerization of the two proteins. This dimerization-based dsDNA sensors will have a range of applications where conventional hybridization-based assay is difficult. |
Volume | 23(8) |
Pages | 1266-71 |
Published | 2008-3-14 |
DOI | 10.1016/j.bios.2007.11.011 |
PII | S0956-5663(07)00478-2 |
PMID | 18178076 |
MeSH | Base Sequence Biosensing Techniques / methods* DNA / chemistry* DNA / genetics* DNA-Binding Proteins / chemistry Dimerization Fluorescence Resonance Energy Transfer / methods* Fluorescent Dyes / chemistry* Luminescent Proteins / chemistry* Molecular Sequence Data |
IF | 10.257 |
Times Cited | 4 |
WOS Category | NANOSCIENCE & NANOTECHNOLOGY CHEMISTRY, ANALYTICAL BIOTECHNOLOGY & APPLIED MICROBIOLOGY ELECTROCHEMISTRY BIOPHYSICS |
Resource | |
DNA material | ETR103 (RDB01198) |