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| T克隆载体的构建 | |
| 引用本文： | 胡学军,徐红梅,赵东利,李晶泉,袁晓东.T克隆载体的构建[J].北京农学院学报,2001,16(4):6-8. |
| 作者姓名： | 胡学军 徐红梅 赵东利 李晶泉 袁晓东 |
| 作者单位： | 1. 大连大学医学院生物教研室, 2. 北京农学院, 3. 宝生物大连有限工程公司, |
| 摘 要： | 本文描述了一种构建与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Ampr 基因的Eam110 5I酶切位点上,以点突变的方式封闭Eam110 5I酶切位点。经转化大肠杆菌JM 10 9证实,该改造过的pUC118质粒,可使宿主细胞具有氨苄抗性,仍可作为氨苄选择标记的克隆载体。将一人工合成的具有两个Eam110 5I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Eam110 5I酶切位点的pUC118 载体的BamHI位点,构成新的克隆载体,此质粒命名为pUC118E。该载体经Eam110 5I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。 |
| 关 键 词： | Tvector pUC118 PCR产物 限制酶Eam1105I |
| 文章编号： | 1002-3186(2001)04-0006-03 |
| 修稿时间： | 2001年6月20日 |
Construction of Cloning Vector for Direct Ligation with PCR Products |
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| Hu Xuejun,Xu Hongmei,Zhao Dongli,Li jingquan,Yuan Xiaodong.Construction of Cloning Vector for Direct Ligation with PCR Products[J].Journal of Beijing Agricultural College,2001,16(4):6-8. | |
| Authors: | Hu Xuejun Xu Hongmei Zhao Dongli Li jingquan Yuan Xiaodong |
| Institution: | Hu Xuejun 2Xu Hongmei 1 Zhao Dongli 3 Li jingquan 3Yuan Xiaodong |
| Abstract: | A new method for construction cloning vector (T-vector)for direct ligation with PCR products was described.The T-vector derived from pUC118 which the unique restriction site of Eam 1105I in the region of Amp r gene was deleted and an artificial DNA fragment flanking two Eam 1105 Iwas introduced at the site of Bam H I.The modified vector was named pUC118E.A T-vector with 3' over hang end of a single T can be obtained via digesting of pUC118E with Eam 1105I.PCR produces can be easy to be cloned with this T-vector. |
| Keywords: | T-vector pUC118 PCR produces Eam 1105I |
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