Position: Home > Articles > Construction of Cloning Vector for Direct Ligation with PCR Products
Journal of Beijing University of Agriculture
2001
(4)
8-10
T克隆载体的构建
作 者：
胡学军;徐红梅;赵东利;李晶泉;袁晓东
单 位：
大连大学医学院生物教研室;北京农学院;宝生物大连有限工程公司
关键词：
Tvector;pUC118;PCR产物;限制酶Eam1105I
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。
译 名：
Construction of Cloning Vector for Direct Ligation with PCR Products
作 者：
Hu Xuejun 2Xu Hongmei 1 Zhao Dongli 3 Li jingquan 3Yuan Xiaodong ( 1) Medical College of Dalian University,Dalain 116622; 2) Bejing Agricultural College,Beijing 102206; 3) Takara Biotechnology Dalian Co.Ltd.,Dalain 116600)
关键词：
T-vector,pUC118,PCR produces, Eam 1105I
摘 要：
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.
导出引用
计量
文章访问数: 12
HTML全文浏览量: 0
PDF下载量: 0
所属期刊
