Zyrina N V, Artiukh R I, Svad'bina I V, Zheleznaia L A, Matvienko N I
Bioorg Khim. 2012 Mar-Apr;38(2):199-205. doi: 10.1134/s1068162012020161.
In the presence of the Nt.BspD6I nicking endonuclease DNA polymerase Bst stimulates intensive template/primer-independent DNA synthesis. Template/primer-independent DNA synthesis could be the reason for appearing nonspecific DNA products in many DNA amplification reactions particularly in the reactions with using nicking endonucleases. Search of the modes for inhibition template/primer-independent DNA synthesis becomes an urgent task because of broadening the DNA amplification methods with using nicking endonucleases. We report here that the E. coli single-stranded DNA binding protein has no effect on the template/primer-independent DNA synthesis. In the absence of the nicking endonuclease the single-stranded DNA binding protein encoded by bacteriophage T4 gene 32 completely inhibits template/primer-independent DNA synthesis. This protein does not inhibit synthesis of specific DNA product in the presence of nicking endonuclease but remarkably decreases the amount of nonspecific products.
在Nt.BspD6I切口内切核酸酶存在的情况下，DNA聚合酶Bst会刺激强烈的模板/引物非依赖性DNA合成。模板/引物非依赖性DNA合成可能是许多DNA扩增反应中出现非特异性DNA产物的原因，特别是在使用切口内切核酸酶的反应中。由于使用切口内切核酸酶拓宽了DNA扩增方法，寻找抑制模板/引物非依赖性DNA合成的模式成为一项紧迫任务。我们在此报告，大肠杆菌单链DNA结合蛋白对模板/引物非依赖性DNA合成没有影响。在没有切口内切核酸酶的情况下，噬菌体T4基因32编码的单链DNA结合蛋白完全抑制模板/引物非依赖性DNA合成。该蛋白在切口内切核酸酶存在的情况下不抑制特异性DNA产物的合成，但显著减少非特异性产物的量。
