Batchikova N V, Skaptsova N V, Tvardovskaia S E, Besidskiĭ E S, Dan'kov Iu V
Bioorg Khim. 1988 May;14(5):621-30.
The solid-phase phosphotriester method was used to synthesise 24 oligodeoxyribonucleotides, which were enzymatically joined together to give the human epidermal growth factor gene and its analogue containing Leu codon in position 21. The primary structure of the cloned genes were confirmed by the Maxam-Gilbert technique. It is demonstrated that the high purity degree of oligonucleotides allows to synthesise and clone genes without purification of intermediate fragments.
采用固相磷酸三酯法合成了24个寡脱氧核糖核苷酸,这些寡核苷酸通过酶促连接在一起,得到人表皮生长因子基因及其在第21位含有亮氨酸密码子的类似物。通过Maxam-Gilbert技术证实了克隆基因的一级结构。结果表明,寡核苷酸的高纯度使得无需纯化中间片段即可合成和克隆基因。