Design of LNA probes that improve mismatch discrimination

被引：259

作者：

You, Yong ^{[1
]}

Moreira, Bernardo G. ^{[1
]}

Behlke, Mark A. ^{[1
]}

Owczarzy, Richard ^{[1
]}

机构：

[1] Integrated DNA Technol, Coralville, IA 52241 USA

来源：

NUCLEIC ACIDS RESEARCH | 2006年 / 34卷 / 08期

关键词：

D O I：

10.1093/nar/gkl175

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Locked nucleic acids (LNA) show remarkable affinity and specificity against native DNA targets. Effects of LNA modifications on mismatch discrimination were studied as a function of sequence context and identity of the mismatch using ultraviolet (UV) melting experiments. A triplet of LNA residues centered on the mismatch was generally found to have the largest discriminatory power. An exception was observed for G-T mismatches, where discrimination decreased when the guanine nucleotide at the mismatch site or even the flanking nucleotides were modified. Fluorescence experiments using 2-aminopurine suggest that LNA modifications enhance base stacking of perfectly matched base pairs and decrease stabilizing stacking interactions of mismatched base pairs. LNAs do not change the amount of counterions (Na+) that are released when duplexes denature. New guidelines are suggested for design of LNA probes, which significantly improve mismatch discrimination in comparison with unmodified DNA probes.

引用

页数：11

共 37 条

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