与三核苷酸共有序列相关的DNA聚合酶停顿的分析与抑制
Analysis and suppression of DNA polymerase pauses associated with a trinucleotide consensus.
作者信息
Mytelka D S, Chamberlin M J
机构信息
Graduate Group in Genetics, University of California, Berkeley 94720, USA.
出版信息
Nucleic Acids Res. 1996 Jul 15;24(14):2774-81. doi: 10.1093/nar/24.14.2774.
Abstract
We have studied a novel class of DNA sequences that cause DNA polymerases to pause. These sequences have the central consensus Py-G-C and are not necessarily adjacent to hairpins in the DNA template. Since most consensus sequences do not cause pauses under standard conditions, additional template features must exist that make it difficult to incorporate nucleotides at these positions. We believe that these pauses result from constraints that make the conformation change involved in nucleotide selection more difficult. These pauses can obscure parts of DNA sequencing ladders and prevent DNA amplification by the polymerase chain reaction. The addition of betaine, and some related compounds, relieves these pauses.
摘要
我们研究了一类能使DNA聚合酶暂停的新型DNA序列。这些序列具有核心共有序列Py-G-C，且不一定与DNA模板中的发夹结构相邻。由于大多数共有序列在标准条件下不会导致暂停，所以必然存在其他模板特征使得在这些位置难以掺入核苷酸。我们认为这些暂停是由一些限制因素导致的，这些因素使得核苷酸选择过程中涉及的构象变化更加困难。这些暂停会模糊DNA测序梯的部分区域，并阻止通过聚合酶链反应进行DNA扩增。添加甜菜碱及一些相关化合物可缓解这些暂停现象。
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