Sung W L, Zahab D M, MacDonald C A, Tam C S
Gene. 1986;47(2-3):261-7. doi: 10.1016/0378-1119(86)90069-7.
A synthetic 'crossover linker' technique has been designed for gene modification. The linker has a restriction end for an initial 'cohesive end' ligation with one terminus of a linearized plasmid, a middle section carrying modified sequence information, and an 'homology-searching' sequence of 20 bp at its other end, that is homologous to a specific region in the opposite terminus of the plasmid. Inside the Escherichia coli transformation host, intramolecular recombination between the homologous ends of the resultant plasmid intermediate completes the integration of the linker. Using different crossover linkers, a human parathyroid hormone gene which had previously been cloned into plasmid pUC8 was converted to mutant coding sequences via specific base substitution, sequence deletion and sequence insertion.
一种用于基因修饰的合成“交叉连接子”技术已被设计出来。该连接子具有一个限制性末端，用于与线性化质粒的一个末端进行初始的“粘性末端”连接，中间部分携带修饰后的序列信息，并且在其另一端有一个20个碱基对的“同源搜索”序列，该序列与质粒相对末端的特定区域同源。在大肠杆菌转化宿主内，所得质粒中间体同源末端之间的分子内重组完成了连接子的整合。使用不同的交叉连接子，先前克隆到质粒pUC8中的人甲状旁腺激素基因通过特定的碱基替换、序列缺失和序列插入被转化为突变编码序列。
