Smith J, Cook E, Fotheringham I, Pheby S, Derbyshire R, Eaton M A, Doel M, Lilley D M, Pardon J F, Patel T, Lewis H, Bell L D
Nucleic Acids Res. 1982 Aug 11;10(15):4467-82. doi: 10.1093/nar/10.15.4467.
A DNA duplex coding for the 53 amino acids of human beta-urogastrone has been synthesised. Computer assisted design of the gene included restriction endonuclease sites for plasmid insertion, a termination codon and two triplets coding for lysine at the 5'-end of the structural gene. The synthesis involved preparation of 23 oligodeoxyribonucleotides by phosphotriester procedures coupled to rapid HPLC techniques. The gene was constructed in two halves by enzymatic ligation of the oligonucleotides and cloned into a specially constructed chimeric plasmid vector. Escherichia coli K12 MRC8 was transformed by the plasmid and clones containing the full gene sequence were isolated and characterised.
已合成了编码人β-尿抑胃素53个氨基酸的DNA双链体。该基因的计算机辅助设计包括用于质粒插入的限制性内切酶位点、一个终止密码子以及在结构基因5′端编码赖氨酸的两个三联体。合成过程涉及通过磷酸三酯法结合快速高效液相色谱技术制备23个寡脱氧核糖核苷酸。通过寡核苷酸的酶促连接将该基因分两半构建，并克隆到一个专门构建的嵌合质粒载体中。用该质粒转化大肠杆菌K12 MRC8，分离并鉴定了含有完整基因序列的克隆。
