Kobets M L, Rivkin M I, Bogachev V S, Kumarev V P
Bioorg Khim. 1988 Jan;14(1):43-7.
A simple and economy method of the biochemical assembling of long double-stranded DNA segments is described. A single-stranded polydeoxynucleotide 122 bases long representing a fragment of synthetic gene of human beta-interferon was assembled from three synthetic fragments 36 (two) and 50 bases long on four complementary 12-mers as templates. This single-stranded polynucleotide was converted, in the presence of DNA polymerase 1 and a 12-meric primer, in to the full-length double-stranded DNA (the beta-interferon gene segment). It was cloned into an E. coli plasmid vector pBR322 and its sequence confirmed.
本文描述了一种简单且经济的长双链DNA片段生化组装方法。一条长度为122个碱基的单链聚脱氧核苷酸，代表人类β-干扰素合成基因的一个片段，它是在四条互补的12聚体作为模板的情况下，由三个分别为36个碱基（两个）和50个碱基长的合成片段组装而成。在DNA聚合酶I和一条12聚体引物存在的情况下，这条单链聚脱氧核苷酸被转化为全长双链DNA（β-干扰素基因片段）。它被克隆到大肠杆菌质粒载体pBR322中，并对其序列进行了确认。
