School of Materials Science and ‡Research Center for Bio-Architecture, Japan Advanced Institute of Science and Technology , 1-1 Asahi-dai, Nomi, Ishikawa 923-1292, Japan.
J Am Chem Soc. 2013 Oct 30;135(43):16161-7. doi: 10.1021/ja406965f. Epub 2013 Oct 22.
To clarify the cis-trans isomeric effect on the ultrafast DNA photo-cross-linking reaction of 3-cyanovinylcarbazole nucleoside ((CNV)K) in the DNA duplex, which gives a single photodimer on the reversed-phase HPLC chromatogram, the kinetics of the cis-trans photoisomerization of (CNV)K in double-stranded DNA was evaluated. Since the photoisomerization rate constant for cis to trans isomerization in double-stranded DNA was significantly larger than that for trans to cis isomerization, and the thermodynamic stability of the trans isomer was higher than that of the cis isomer, it was strongly suggested that the trans isomer of (CNV)K is a reactive species of the photo-cross-linking reaction. (1)H-(1)H NOESY analysis of the photoadduct consisting of (CNV)K and T also supported the trans-mediated photo-cross-linking reaction of (CNV)K. By using this ultrafast photo-cross-linking reaction for the molecular beacon-based SNPs typing, four individual Japanese rice strains were clearly distinguishable with simple photoirradiation and fluorescence imaging using double-stranded target DNAs.
为了阐明顺反异构效应对 3-氰基乙烯基咔唑核苷（（CNV）K）在 DNA 双链体中超快光交联反应的影响，该反应在反相 HPLC 色谱图上给出单个光二聚体，评估了（CNV）K 在双链 DNA 中的顺反异构光异构化的动力学。由于双链 DNA 中顺式到反式异构化的光异构化速率常数明显大于反式到顺式异构化的光异构化速率常数，并且反式异构体的热力学稳定性高于顺式异构体，因此强烈表明（CNV）K 的反式异构体是光交联反应的反应性物质。（1）H-（1）H NOESY 分析由（CNV）K 和 T 组成的光加合物也支持（CNV）K 的反式介导光交联反应。通过使用这种超快光交联反应进行基于分子信标的 SNPs 分型，使用双链靶 DNA 进行简单的光辐照和荧光成像，可清楚地区分四个个体日本水稻品系。
