W. A. Engelhardt Institute of Molecular Biology Russian Academy of Sciences, Vavilov St. 32, Moscow, Russia, 119991.
Sci Rep. 2021 Jan 28;11(1):2423. doi: 10.1038/s41598-021-82150-2.
Replicative strand slippage is a biological phenomenon, ubiquitous among different organisms. However, slippage events are also relevant to non-natural replication models utilizing synthetic polymerase substrates. Strand slippage may notably affect the outcome of the primer extension reaction with repetitive templates in the presence of non-natural nucleoside triphosphates. In the current paper, we studied the ability of Taq, Vent (exo-), and Deep Vent (exo-) polymerases to produce truncated, full size, or expanded modified strands utilizing non-natural 2'-deoxyuridine nucleotide analogues and different variants of the homopolymer template. Our data suggest that the slippage of the primer strand is dependent on the duplex fluttering, incorporation efficiency for a particular polymerase-dNTP pair, rate of non-templated base addition, and presence of competing nucleotides.
复制链滑动是一种普遍存在于不同生物体中的生物学现象。然而，滑动事件也与利用合成聚合酶底物的非自然复制模型有关。在存在非天然核苷三磷酸的情况下，链滑动可能会显著影响重复模板的引物延伸反应的结果。在本文中，我们研究了 Taq、Vent（外切）和 Deep Vent（外切）聚合酶利用非天然 2'-脱氧尿苷核苷酸类似物和不同的同源聚合物模板变体产生截断、全长或扩展修饰链的能力。我们的数据表明，引物链的滑动取决于双链的摆动、特定聚合酶-dNTP 对的掺入效率、非模板碱基添加的速率以及竞争核苷酸的存在。
