一种用于荧光检测三链螺旋DNA的光纤生物传感器。
A fiber optic biosensor for fluorimetric detection of triple-helical DNA.
作者信息
Uddin A H, Piunno P A, Hudson R H, Damha M J, Krull U J
机构信息
Department of Chemistry, Otto Maas Chemistry Building, McGill University, Montreal, Quebec H3A 2K6, Canada.
出版信息
Nucleic Acids Res. 1997 Oct 15;25(20):4139-46. doi: 10.1093/nar/25.20.4139.
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 oligonucleotide, dT10, 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 triplex, 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.
一种光纤生物传感器用于荧光检测T/AT三链螺旋DNA的形成。两组熔融石英光纤的表面用六亚乙基氧化物连接基进行功能化处理，然后使用DNA合成仪分别在3'至5'和5'至3'方向上生长十聚腺苷酸寡核苷酸。杂交的荧光研究表明，固定在光纤上的寡聚物与溶液相中的互补寡核苷酸之间发生了明确的杂交，这可通过嵌入的溴化乙锭发出的荧光检测到。互补寡核苷酸dT10在将系统冷却至双链体解链温度（Tm）以下时预期会发生沃森-克里克杂交，其荧光强度具有负温度系数。进一步冷却至嘧啶基序T*AT三链螺旋形成的温度点时，观察到荧光强度变化具有正温度系数。已知与分支核酸形成的反向-霍格施泰因T·AT三链体随着温度降低荧光强度相应降低。几分钟内即可实现完整的分析信号演变。
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