Design of LNA probes that improve mismatch discrimination.
Design of LNA probes that improve mismatch discrimination.
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LNA探针的设计可改善不匹配歧视。
DOI：
10.1093/nar/gkl175
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发表时间：
2006-05-02
影响因子：
14.9
通讯作者：
Owczarzy, Richard
中科院分区：
文献类型：
--
作者：
You, Yong;Moreira, Bernardo G.;Behlke, Mark A.;Owczarzy, Richard
锁定核酸(LNA)对天然DNA靶标具有显著的亲和力和特异性。利用紫外(UV)熔融实验研究了LNA修饰对错配识别的影响，作为序列上下文和错配识别的函数。以错配为中心的三联体LNA残基通常被发现具有最大的区分力。G-T错配是一个例外，当错配位置的鸟嘌呤核苷酸甚至侧翼核苷酸被修饰时，辨别力降低。使用2-氨基嘌呤的荧光实验表明，LNA修饰增强了完全匹配碱基对的碱基堆积，降低了不匹配碱基对的稳定堆积作用。RNA不会改变双链变性时释放的反离子(Na+)的量。建议了新的LNA探针设计指南，与未修饰的DNA探针相比，该指南显著改善了错配识别能力。
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.
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