Fluorosensor proteins to detect specific DNA sequences in living bacteria.
While there are many hybridization-based DNA sensors, few of them can detect native double-stranded DNA, which is most commonly found in physiological conditions. Here we made novel fluorosensor proteins comprised of a pair of two zinc fingers tethered with an N-terminal dimerization motif and a C-terminal yellow fluorescent protein fragment (split eYFP) to detect specific DNA sequence in a living bacteria. When E. coli Top10 cells harboring the plasmid encoding the fusion proteins and a test plasmid encoding target DNA sequence were induced for the protein expression, significant increase in fluorescence was observed, compared with the strain harboring a test plasmid without target DNA sequence.
Waki
Shoko
S
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Japan.
Yoshitake
Kazutoshi
K
Iwasaki
Ryohei
R
Ueda
Hiroshi
H
D001426:Bacterial Proteins / Q000737:chemistry*; D019281:Dimerization; D005456:Fluorescent Dyes / Q000737:chemistry*; D008164:Luminescent Proteins / Q000737:chemistry*; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA / Q000379:methods*; D013050:Spectrometry, Fluorescence; D016335:Zinc Fingers
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria