Drutsa V L, Bednarek P Z, Koroleva O N
Bioorg Khim. 1994 Nov;20(11):1206-17.
The in vitro replication of synthetic oligodeoxyribonucleotides carrying internucleotide polyphosphate groups or alkanediol "spacers" of various sizes with the use of various DNA polymerases has been studied. All modifications, except for the diphosphate group, almost completely block the polymerization process. In the case of AMV reverse transcriptase, Taq and T7 DNA polymerases and also the Klenow fragment of E. coli DNA polymerase I, a template-independent addition of a nucleotide at the 3' end of the incomplete replica was observed. T4 DNA polymerase, displaying the strongest 3'-5' exonuclease activity among the polymerases studied, did not incorporate additional nucleotides. The use of oligonucleotides with non-nucleotide inserts as primers in polymerase chain reaction (PCR) allows to obtain DNA copies with protruding 5'-termini, suitable for hybridisation analysis.
研究了使用各种DNA聚合酶对携带核苷酸间多磷酸基团或不同大小链烷二醇“间隔物”的合成寡脱氧核糖核苷酸进行体外复制的情况。除了二磷酸基团外，所有修饰几乎完全阻断了聚合过程。对于禽成髓细胞瘤病毒（AMV）逆转录酶、Taq和T7 DNA聚合酶以及大肠杆菌DNA聚合酶I的Klenow片段，在不完全复制产物的3'末端观察到了与模板无关的核苷酸添加。在所研究的聚合酶中，T4 DNA聚合酶具有最强的3'-5'核酸外切酶活性，它不会掺入额外的核苷酸。在聚合酶链反应（PCR）中使用带有非核苷酸插入物的寡核苷酸作为引物，可以获得具有突出5'末端的DNA拷贝，适用于杂交分析。
