Inhibition of multiple thermostable DNA polymerases by a heterodimeric aptamer

被引:53
作者
Lin, Y [1 ]
Jayasena, SD [1 ]
机构
[1] NEXSTAR PHARMACEUT INC,BOULDER,CO 80301
关键词
Taq DNA polymerase; Stoffel fragment; Tth DNA polymerase affinity selection; ''hot start'' PCR;
D O I
10.1006/jmbi.1997.1165
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
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. (C) 1997 Academic Press Limited.
引用
收藏
页码:100 / 111
页数:12
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