Fujimoto Kenzo, Toyosato Kei, Nakamura Shigetaka, Sakamoto Takashi
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahi-dai, Nomi, Ishikawa 923-1292, Japan.
Bioorg Med Chem Lett. 2016 Nov 1;26(21):5312-5314. doi: 10.1016/j.bmcl.2016.09.035. Epub 2016 Sep 15.
Photo-cross-linkable fluorescent oligodeoxyribonucleotides having 3-cyanovinylcarbazole nucleoside were applied to fluorescence in situ hybridization (FISH) based 16S rRNA detection in Escherichia coli cells. As the photo-cross-linked probe/rRNA hybrid was stable under the denaturing condition, decrease of the fluorescence signal through the washing process was prevented. The thermally irreversible hybridization property also enabled stable hybridization with the structured region on the target RNA, and facilitated design of the sequence for the FISH probe. Further development of the method might contribute to quantitative and stable FISH staining.
含有3-氰基乙烯基咔唑核苷的可光交联荧光寡脱氧核糖核苷酸被应用于基于荧光原位杂交（FISH）的大肠杆菌细胞16S rRNA检测。由于光交联的探针/rRNA杂交体在变性条件下稳定，因此可防止洗涤过程中荧光信号的降低。热不可逆杂交特性还能与靶RNA的结构区域稳定杂交，并有助于设计FISH探针的序列。该方法的进一步发展可能有助于实现定量且稳定的FISH染色。
