摘要： 本文描述一种构建与PCR产物直接连接的克隆载体的方法。以高拷贝克隆载体pUC118为骨架载体，在pUC118质粒氨苄抗性基因的Eam1105 I 酶切位点上，以点突变的方式封闭Eam1105 I 酶切位点。经转化大肠杆菌JM109证实，该改造过的pUC118质粒，可使宿主细胞仍具有氨苄抗性。将一人工合成的具有两个Eam1105 I 酶切位点的互补寡聚核苷酸链（两端具有BamH I 接头）插入已封闭Eam1105 I 酶切位点的pUC118*载体的BamH I 位点，构成新的克隆载体，此质粒命名为pUC118E。该载体经Eam1105 I 酶切后，可产生3′末端突出一个T碱基的T-载体，能与PCR产物直接连接。
Abstract:A new method for construction of a cloning vector (T-vector) for direct ligation with PCR products was described.The T-vector derived from pUC118 in which the unique restriction site of Eam1105 I in the region of Ampr gene was deleted and an artificial DNA fragment flanking two Eam1105 I was introduced at the site of BamHI.The modified vector was named as pUC118E.A T-vector with 3′over hang end of a single T can be obtained via digesting of pUC118E with Eam1105I.PCR products can be easily cloned with this T-vector.