Sung W L, Zahab D M, Yao F L, Wu R, Narang S A
Nucleic Acids Res. 1986 Aug 11;14(15):6159-68. doi: 10.1093/nar/14.15.6159.
Simultaneous synthesis of two DNA duplexes encoding human and mouse epidermal growth factors (EGF) was accomplished in a single step. A 174 b.p. DNA heteroduplex, with 16 single and double base pair mismatches, was designed. One strand encoded the human EGF, and the opposite strand indirectly encoded the mouse EGF. The heteroduplex DNA was synthesized by ligation of seven overlapping oligodeoxyribonucleotides with a linearized plasmid. After transformation in E. coli HB101 (recA 13), the resulting heteroduplex plasmid served as the template in plasmid replication. Two different plasmid progenies bearing either the human or mouse EGF-coding sequence were identified by colony hybridization using the appropriate probes. However, in E. coli JM103, the same process yielded plasmid progenies encoding different chimeric EGF molecules, presumably due to crossover of human and mouse EGF gene sequences.
在一步反应中同时完成了编码人表皮生长因子（EGF）和小鼠表皮生长因子的两条DNA双链体的合成。设计了一个174碱基对的DNA异源双链体，其含有16个单碱基对和双碱基对错配。一条链编码人EGF，相对的链间接编码小鼠EGF。通过将七个重叠的寡脱氧核糖核苷酸与线性化质粒连接来合成异源双链体DNA。在大肠杆菌HB101（recA 13）中进行转化后，所得的异源双链体质粒在质粒复制中用作模板。使用合适的探针通过菌落杂交鉴定出携带人或小鼠EGF编码序列的两种不同的质粒后代。然而，在大肠杆菌JM103中，相同的过程产生了编码不同嵌合EGF分子的质粒后代，推测这是由于人和小鼠EGF基因序列的交叉。
