Molecular beacons for isothermal fluorescence enhancement by the cleavage of RNase HII from Chlamydia pneumoniae.
This article describes a new assay for isothermal enhancement of fluorescence intensity. The assay is based on the cleavage of duplexes formed by the chimeric DNA-rN(1)-DNA molecular beacon (cMB) and target DNA with Chlamydia pneumoniae RNase HII (CpRNase HII). The loop sequence of the cMB, which was designed according to the target sequence, contains a single ribonucleotide. The combination of CpRNase HII cleavage and cMB (RHMB) permitted a 90-fold increase in fluorescence intensity change compared with the hybridization reaction in the presence of the same amount of target DNA. These results indicate that the RHMB assay can enhance the fluorescence signal in real-time monitoring of the target DNA.
Hou
Jingli
J
College of Life Science and Technology, Shanghai Jiao Tong University, Shanghai 200030, China.
Liu
Xipeng
X
Wang
Jian
J
Liu
Jianhua
J
Duan
Tao
T
D016993:Chlamydophila pneumoniae / Q000201:enzymology*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D015345:Oligonucleotide Probes / Q000737:chemistry*; D016914:Ribonuclease H / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013696:Temperature; D013997:Time Factors
D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D004247:DNA; C091139:ribonuclease HII; D016914:Ribonuclease H