| T克隆载体的构建 |
| |
| 引用本文： | 胡学军,李江,袁晓东,李晶泉,安利佳.T克隆载体的构建[J].大连大学学报,1998(6). |
| |
| 作者姓名： | 胡学军 李江 袁晓东 李晶泉 安利佳 |
| |
| 作者单位： | 医学院生物教研室，辽宁师范大学生命科学院，宝生物大连有限工程公司，宝生物大连有限工程公司，辽宁师范大学生命科学院 大连 116622，116600，116600 |
| |
| 摘 要： | 以高拷贝克隆载体pUC118为骨架载体,在pUC118质粒Amp~r基因的Eam11057酶切位点上,以点突变的方式封闭Eam11051酶切位点,将一人工合成的具有两个Eam11051酶切位点的互补寡聚核苷酸链(两端具有BamHI接头)插入已封闭Eam1105I酶切位点的pUC118质粒的BamHI位点,构成新的克隆载体,此质粒命名为pUC118E,该载体经Eam11051酶切后,可产生3’端突出一个T碱基的T-vector,能与PCR产物直接连接,经转化大肠杆菌JM109证实,该改造过的pUC118质粒,仍可使宿主细胞具有氨苄抗性,可作为氨苄选择标记的克隆载体。
|
| 关 键 词： | T-vector pUC118 PCR产物 Eam11051 |
Construction of Cloning Vector for Direct Ligation with PCR Products |
| |
| Hu Xuejun Li Jiang Yuan Xiaodong Li Jingquan An Lijia.Construction of Cloning Vector for Direct Ligation with PCR Products[J].Journal of Dalian University,1998(6). |
| |
| Authors: | Hu Xuejun Li Jiang Yuan Xiaodong Li Jingquan An Lijia |
| |
| Institution: | Hu Xuejun Li Jiang Yuan Xiaodong Li Jingquan An Lijia(Medical Department of Dalian University) (Bioscience College of Liaoning Normal University) (TaKaRa Dalian Bioengineering Co. Ltd.) |
| |
| Abstract: | Construction of A New Type of T-vector A new type of T-vector derived from pUC118 cloning vector was abtained. The unique restriction site of Eam1105I in the Amp gene was deleted, and an artificial DNA fragment flanking two new Eam1105I sites was introduced at BamHI site. The modified plasmid was named pUCHSE. A T-vector with 3'over hang end of a single T can be obtained by digesting of pUC118E using Eam1105I, and the T-vector can be directly ligated with PCR products. It was proved that the E.coli JM109 host cell transformed with pUC118E has the ability of Amp resistance. |
| |
| Keywords: | T-vector pUC118 PCR products Eam1105I |
| 本文献已被 CNKI 等数据库收录！ |
