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Tth DNA聚合酶基因的分段PCR克隆及其表达载体构建
引用本文:蒋永强,蒙健宗,黄鲲,黄日波.Tth DNA聚合酶基因的分段PCR克隆及其表达载体构建[J].广西农业生物科学,1999,18(1):33-38.
作者姓名:蒋永强  蒙健宗  黄鲲  黄日波
作者单位:广西大学生物技术实验中心,南宁,530005
基金项目:广西自然科学基金;桂科青9418011;
摘    要:利用计算机软件对取自GenBank的嗜热栖热菌ThermusthermophilusHB-8的DNA聚合酶基因(Tth)进行分析,并设计了两组引物进行PCR,分段克隆该基因。将两个PCR产物同时连接到另一个克隆载体上,使之成为完整基因,最后将完整基因克隆到表达载体上,构建了Tth基因的表达质粒,使之适合在以大肠杆菌为寄主的细胞中进行表达。

关 键 词:嗜热栖热菌  DNA聚合酶  Tth基因  分段PCR  克隆
修稿时间:1999-01-16

SegmentedPCR cloning of Tth DNA polymerase Gene and Construction of its Expression Vector
Jiang Yongqiang,Meng Jianzhong,Huang Kun,Huang Ribo.SegmentedPCR cloning of Tth DNA polymerase Gene and Construction of its Expression Vector[J].Journal of Guangxi Agricultural and Biological Science,1999,18(1):33-38.
Authors:Jiang Yongqiang  Meng Jianzhong  Huang Kun  Huang Ribo
Abstract:The sequence of a DNA polymerase gene (Tth) from extremthermophile Thermus thermophilus HB-8 was obtained from GenBank and analysed using PC-based DNA-analysing program. Two pairs of PCR primers were designed in order to clone two segments of Tth gene separately. Two segments were joined in a cloning plasmid to from intact Tth gene and then subcloned onto an expression vector suitable for being expressed in E. coli host cells.
Keywords:Thermus thermophilus  DNA polymerase  Tth  gene  segmented-PCR  cloning  
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