Design of Environmentally Sensitive Fluorescent Nucleosides and Their Application to Innovative Fluorescence Probe for Gene Analysis

我们设计了一种新型的碱基区分荧光(BDF)核苷，用于SNP基因分型和基因检测。我们还成功地设计了环境敏感的荧光核苷，根据环境敏感核苷基座附近的局部环境的不同，在不同的波长发射出强烈的荧光。新设计的ESF核苷可以通过发射波长及其强度的显著变化来区分结构变化，如单链、匹配、错配和缺失。利用新设计的ESF核苷首次发现了猝灭分子信标和末端自由分子信标。这些低成本的MB被用于细胞中的基因检测。还设计了对pH敏感的ESF核苷。这种pH传感器ESF核苷只有在pH 5.5以下才能发出强烈的荧光，用于癌细胞的检测。

We have designed a novel type of base-discriminating fluorescent (BDF) nucleosides which are used for SNP genotyping and gene detection. We also succeeded to design environmentally sensitive fluorescent (ESF) nucleosides that emit strong fluorescence at different wavelengths depending upon a difference of local environment near the ESF base. The newly designed ESF nucleosides may discriminate structural changes such as single strands, matches, mismatches and deletions when incorporated in DNA by a significant change of emission wavelengths as well as their intensities. Quencher and end free molecular beacons (MB) were discovered for the first time by using newly designed ESF nucleosides. These low cost MB were used for the gene detection in a cell. ESF nucleosides that are sensitive to pH were also designed. Such pH sensor ESF nucleoside that emits strong fluorescence only at bellow pH 5.5 was used for the detection of cancer cell.