Lin Y, Jayasena S D
NeXstar Pharmaceuticals Inc., Boulder, CO 80301, USA.
J Mol Biol. 1997 Aug 8;271(1):100-11. doi: 10.1006/jmbi.1997.1165.
Single-stranded DNA aptamers that recognize DNA polymerase from Thermus acquaticus (Taq pol) with high affinity have been described recently. These aptamers have been shown to efficiently inhibit the polymerase activity of Taq pol and are useful in enhancing the amplification efficiency of low copy number targets by the polymerase chain reaction (PCR). Aptamers selected to bind to Taq pol fell into two different sequence families and inhibited several DNA polymerases isolated from the Thermus species, including that from Thermus thermophilus (Tth pol). Aptamers from one sequence family inhibited the Stoffel fragment of Taq pol efficiently, whereas those from the other family did not. Truncated aptamers derived from two parent ligands from both families were combined to form a heterodimeric aptamer that effectively inhibited all three polymerases and were shown to be useful in detecting a low copy number target by PCR amplification. These data demonstrate that the combination of aptamers with different properties into a single molecule broadens their spectrum of utility.
最近已报道了能以高亲和力识别嗜热水生栖热菌（Taq pol）DNA聚合酶的单链DNA适配体。这些适配体已被证明能有效抑制Taq pol的聚合酶活性，并且在通过聚合酶链反应（PCR）提高低拷贝数靶标的扩增效率方面很有用。筛选出的与Taq pol结合的适配体分为两个不同的序列家族，并能抑制从栖热菌属分离出的几种DNA聚合酶，包括嗜热栖热菌（Tth pol）的DNA聚合酶。来自一个序列家族的适配体能有效抑制Taq pol的Stoffel片段，而来自另一个家族的则不能。将源自两个家族的两种亲本配体的截短适配体组合形成异二聚体适配体，该异二聚体适配体能有效抑制所有三种聚合酶，并被证明可用于通过PCR扩增检测低拷贝数靶标。这些数据表明，将具有不同特性的适配体组合成单个分子可拓宽其应用范围。
