Ota N, Hirano K, Warashina M, Andrus A, Mullah B, Hatanaka K, Taira K
National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology, MITI, Tsukuba Science City, Japan.
Nucleic Acids Symp Ser. 1997(37):207-8.
We examined changes in the extent of fluorescence resonance energy transfer (FRET) between two different fluorochromes attached to a single oligonucleotide in the presence or absence of target nucleic acids with a specific sequence and a higher-ordered structure. In our system, FRET was maximal when probes were free in solution and a decrease in FRET was evidence of successful hybridization. Incubation of the probe with a single-stranded complementary oligonucleotide reduced the FRET. While, a small change in FRET was also observed when the probe was incubated with an oligonucleotide in which the target site had been embedded in a stable hairpin structure. These results indicate that this spectrofluorometric method and FRET probes can be used to estimate the efficacy of hybridization between a probe and its target site within highly ordered structures. It should help us to estimate the suitability of designed functional molecules, such as antisense DNA and RNA and ribozymes, that target to specific sites.
我们研究了在存在或不存在具有特定序列和高级结构的靶核酸的情况下，连接到单个寡核苷酸上的两种不同荧光染料之间荧光共振能量转移（FRET）程度的变化。在我们的系统中，当探针在溶液中游离时FRET最大，FRET的降低是成功杂交的证据。将探针与单链互补寡核苷酸孵育会降低FRET。同时，当探针与靶位点已嵌入稳定发夹结构的寡核苷酸孵育时，也观察到FRET有微小变化。这些结果表明，这种荧光光谱法和FRET探针可用于评估探针与其在高度有序结构内的靶位点之间杂交的效率。它应该有助于我们评估设计的功能分子的适用性，例如靶向特定位点的反义DNA、RNA和核酶。
