A Label-free Time-resolved Luminescent Platform for Sensitive Endonuclease V Detection Based on Exonuclease III Regulated DNA-Tb3+Luminescence.
Endonuclease V (EndoV) plays the important role of nucleotide excision repair (NER) in the maintenance of genomic stability. Highly sensitive detection of EndoV was achieved through an oligonucleotides sensitizing Tb3+luminescent technique. We found that although both guanine-rich (G-rich) single-stranded DNA and dGMP could enhance the time-resolved luminescence of Tb3+, their efficiencies of enhancement were considerably different. Employing such interesting phenomenon, a label-free and time-resolved luminescent strategy for the sensitive detection of EndoV activity was developed based on DNA-enhanced time-resolved luminescence (TRL) of Tb3+. The EndoV was used to cut off the deoxyinosine site (dI) and convert the 3'-protruding termini to a recessed end, and Exonuclease III (Exo III) was used to enhance the signal contrast via digestion of G-rich DNA to dNTP. Combining with the natural advantages of the TRL, the proposed method exhibited a good linear response to EndoV ranging from 0.005 to 0.4 U/mL, with a low limit of detection of 0.005 U/mL.
State Environmental Protection Key Laboratory of Drinking Water Source Management and Technology, Shenzhen Key Laboratory of Drinking Water Source Safety Control, Shenzhen Research Academy of Environmental Sciences.
Huaijun
H
Nie
Huifen
H
Huang
Wang
W
Li
Tao
T
Yang
D015374:Biosensing Techniques / Q000379:methods; D004277:DNA, Single-Stranded / Q000737:chemistry*; D043224:Deoxyribonuclease (Pyrimidine Dimer) / Q000737:chemistry*; D005090:Exodeoxyribonucleases / Q000737:chemistry*; D057230:Limit of Detection; D049449:Luminescence; D049408:Luminescent Agents; D008163:Luminescent Measurements / Q000379:methods; D013725:Terbium / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry*
D004277:DNA, Single-Stranded; D049408:Luminescent Agents; D014764:Viral Proteins; D013725:Terbium; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III; C470698:endonuclease V, phage T4; D043224:Deoxyribonuclease (Pyrimidine Dimer)