College of Life Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou, Zhejiang, 310058, P.R. China.
Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, P.R. China.
Sci Rep. 2018 Jan 31;8(1):1990. doi: 10.1038/s41598-018-20127-4.
The proofreading activity of the archaeal family B DNA polymerases enables PCR with high fidelity. However, thermostable proofreading DNA polymerases occasionally failed to amplify target fragment that could be amplified by Taq DNA polymerase. We have previously showed that G-rich sequences, which form G-quadruplex, can bind to and inhibit proofreading DNA polymerases. Here we showed that single-stranded oligonucleotides containing sequences of TT(N)mGCCTC can bind and inhibit archaeal family B DNA polymerases but not Taq DNA polymerase. It is very likely that TT(N)mGCCTC inhibits thermostable DNA polymerases during PCR in a single-stranded form. To the best of our knowledge, this is the first example of DNA sequence that could inhibit DNA polymerase in its single-stranded form.
古菌 B 家族 DNA 聚合酶的校对活性使 PCR 具有高保真度。然而，热稳定的校对 DNA 聚合酶偶尔无法扩增可被 Taq DNA 聚合酶扩增的靶片段。我们之前已经表明，富含 G 的序列形成 G-四链体，可与校对 DNA 聚合酶结合并抑制其活性。在此，我们表明含有 TT（N）mGCCTC 序列的单链寡核苷酸可与古菌 B 家族 DNA 聚合酶结合并抑制其活性，但不与 Taq DNA 聚合酶结合。在 PCR 过程中，TT（N）mGCCTC 很可能以单链形式抑制热稳定 DNA 聚合酶。据我们所知，这是第一个可以在单链形式下抑制 DNA 聚合酶的 DNA 序列的例子。
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