Highly Selective Single Nucleotide Polymorphism Recogniton by a Chiral (5S) PNA Beacon

被引:20
作者
Totsingan, Filbert [1 ]
Tedeschi, Tullia [1 ]
Sforza, Stefano [1 ]
Corradini, Roberto [1 ]
Marchelli, Rosangela [1 ]
机构
[1] Univ Parma, Dipartimento Chim Organ & Ind, I-43100 Parma, Italy
关键词
PNA beacons; DNA recognition; chiral monomers; IE-HPLC; fluorescence detection; single nucleotide polymorphisms; PEPTIDE NUCLEIC-ACID; EXCHANGE CHROMATOGRAPHIC ANALYSIS; GENETICALLY-MODIFIED ORGANISMS; LIGHT-UP PROBES; MOLECULAR BEACONS; DNA-BINDING; MICROARRAY TECHNOLOGY; MISMATCH RECOGNITION; STRAND DISPLACEMENT; CYSTIC-FIBROSIS;
D O I
10.1002/chir.20659
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
A chiral peptide nucleic acid (PNA) beacon containing a C-5 modified monomer based on L-lysine was synthesized. The terminal amino group of the lysine side chain was linked to a spacer for future applications on surfaces. The PNA beacon bears a carboxyfluorescein fluorophore and a dabcyl quencher at opposite ends. The DNA binding properties were compared with those of a homologous PNA beacon containing only achiral monomers. Both beacons underwent a fluorescence increase in the presence of complementary DNA, with higher efficiency and higher selectivity (evaluated using single mismatched DNA sequences) observed for the chiral monomer containing PNA. Ion exchange (IE) HPLC with fluorimettic detection was used in combination with the beacon for the selective detection of complementary DNA. A fluorescent peak corresponding to the PNA beacon:DNA duplex was observed at a very low detection limit (1 nM). The discriminating capacity of the chiral PNA beacon for a single mismatch was found to be superior to those observed with the unmodified one, thus confirming the potency of chirality for increasing the affinity and specificity of DNA recognition. Chirality 21:245-253, 2009. (c) 2008 Wiley-Liss, Inc.
引用
收藏
页码:245 / 253
页数:9
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