Prokhorenko Igor A, Astakhova Irina V, Momynaliev Kuvat T, Zatsepin Timofei S, Korshun Vladimir A
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya, Moscow, Russia.
Methods Mol Biol. 2009;578:209-22. doi: 10.1007/978-1-60327-411-1_13.
Excimer formation is a unique feature of some fluorescent dyes (e.g., pyrene) which can be used for probing the proximity of biomolecules. Pyrene excimer fluorescence has previously been used for homogeneous detection of single nucleotide polymorphism (SNP) on DNA. 1-Phenylethynylpyrene (1-1-PEPy), a photostable pyrene derivative with redshifted fluorescence, is able to form excimers (emission maximum about 500-510 nm) and is well suitable for nucleic acid labeling. We have shown the utility of 1-1-PEPy in the excimer-forming DNA probes for detection of 2144A/G and 2143A/G transitions, and 2143A/C substitution in the 23S ribosomal RNA gene of Helicobacter pylori strains resistant to clarithromycin. The phenylethynylpyrene pair can be generated either from 1-1-PEPy pseudonucleoside 4-[4-(pyren-1-ylethynyl)phenyl]-1,3-butanediol or from 2'-O-(1-PEPy) modified nucleosides--2'-O-[3-(pyren-1-ylethynyl)benzyl]uridine and 2'-O-[4-(pyren-1-ylethynyl)benzyl]uridine.
准分子形成是一些荧光染料（如芘）的独特特性，可用于探测生物分子的接近程度。芘准分子荧光此前已用于DNA上单核苷酸多态性（SNP）的均相检测。1-苯基乙炔基芘（1-1-PEPy）是一种具有红移荧光的光稳定芘衍生物，能够形成准分子（发射最大值约为500-510nm），非常适合用于核酸标记。我们已经证明了1-1-PEPy在形成准分子的DNA探针中用于检测对克拉霉素耐药的幽门螺杆菌菌株23S核糖体RNA基因中的2144A/G和2143A/G转换以及2143A/C替换的效用。苯基乙炔基芘对可以由1-1-PEPy假核苷4-[4-(芘-1-基乙炔基)phenyl]-1,3-丁二醇或由2'-O-(1-PEPy)修饰的核苷——2'-O-[3-(芘-1-基乙炔基)苄基]尿苷和2'-O-[4-(芘-1-基乙炔基)苄基]尿苷产生。
