| Tth DNA聚合酶基因的分段PCR克隆及其表达载体构建 |
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| 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 |
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| 作者姓名： | 蒋永强 蒙健宗 黄鲲 黄日波 |
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| 作者单位： | 广西大学生物技术实验中心,南宁,530005 |
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| 基金项目： | 广西自然科学基金;桂科青9418011; |
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| 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。
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| 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 |
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| 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 |
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| Authors: | Jiang Yongqiang Meng Jianzhong Huang Kun Huang Ribo |
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| 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. |
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| Keywords: | Thermus thermophilus DNA polymerase Tth gene segmented-PCR cloning |
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