Published on Sept. 1, 2005 in Nucleic Acids Symp Ser (Oxf) volume (49):285-6.
PubMed ID: 17150745
Nucleoside derivatives of 2-amino-6-(2-thienyl)purine (s) are fluorescent and can be site-specifically incorporated into RNA by transcription mediated by unnatural base pairs between s and its complementary bases. To utilize the fluorescent s base as a probe, we examined the fluorescent properties of s in DNA and RNA fragments. The nucleoside of s exhibited a fluorescence emission centered at 432 nm, characterized by two major excitation maxima (299 and 352 nm), and its quantum yield was 0.41 at pH 7.0. The fluorescence intensity of s in DNA fragments (12-mer) differed depending on circumstances such as the base components, stacking, and pairings. The s base was introduced into a tRNA at specific positions by specific transcription using the unnatural base pairs. The fluorescence intensity of s at each position reflected the local structural features of the tRNA molecules.