A fiber optic biosensor for fluorimetric detection of triple-helical DNA

被引:47
作者
Uddin, AH
Piunno, PAE
Hudson, RHE
Damha, MJ
Krull, UJ
机构
[1] MCGILL UNIV,DEPT CHEM,MONTREAL,PQ H3A 2K6,CANADA
[2] UNIV TORONTO,ERINDALE COLL,DEPT CHEM,CHEM SENSORS GRP,MISSISSAUGA,ON L5L 1C6,CANADA
基金
加拿大自然科学与工程研究理事会;
关键词
D O I
10.1093/nar/25.20.4139
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A fiber optic biosensor was used for the fluorimetric detection of T/AT triple-helical DNA formation, The surfaces of two sets of fused silica optical fibers were functionalized with hexaethylene oxide linkers from which decaadenylic acid oligonucleotides were grown in the 3' to 5' and 5' to 3' direction, respectively, using a DNA synthesizer, Fluorescence studies of hybridization showed unequivocal hybridization between oligomers immobilized on the fibers and complementary oligonucleotides from the solution phase, as detected by fluorescence from intercalated ethidium bromide, The complementary origonucleotide, dT(10), which was expected to Watson-Crick hybridize upon cooling the system below the duplex melting temperature (T-m), provided a fluorescence intensity with a negative temperature coefficient, Upon further cooling, to the point where the pyrimidine motif T*AT triple-helix formation occurred, a fluorescence intensity change with a positive temperature coefficient was observed, The reverse-Hoogsteen. T.AT tripler, which is known to form with branched nucleic acids, provided a corresponding decrease in fluorescence intensity with decreasing temperature, Full analytical signal evolution was attainable in minutes.
引用
收藏
页码:4139 / 4146
页数:8
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