Phenylethynylpyrene excimer forming hybridization probes for fluorescence SNP detection.

Igor A. Prokhorenko*, Irina V. Astakhova, Kuvat T. Momynaliev, Timofei S. Zatsepin, Vladimir A. Korshun, Irina Kira Astakhova

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Abstrakt

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.

OriginalsprogEngelsk
TidsskriftMethods in Molecular Biology
Vol/bind578
Sider (fra-til)209-222
Antal sider14
ISSN1064-3745
StatusUdgivet - 2009

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