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| 一种构建新型T载体的简便方法及应用 | |
| 引用本文： | 齐向辉,陈辉,沈琦,何晨曦,刘学,郭齐,陈华友,徐虹. 一种构建新型T载体的简便方法及应用[J]. 生物技术通报, 2011, 0(3) |
| 作者姓名： | 齐向辉 陈辉 沈琦 何晨曦 刘学 郭齐 陈华友 徐虹 |
| 作者单位： | 1. 江苏大学食品与生物工程学院,镇江,212013;南京工业大学材料化学工程国家重点实验室,南京,210009 2. 江苏大学食品与生物工程学院,镇江,212013 3. 南京工业大学材料化学工程国家重点实验室,南京,210009 |
| 基金项目： | 国家自然科学基金项目(21006041); 江苏省博士后基金项目(0901012B); 江苏大学高级专业人才科研资助项目(08JDG009),江苏大学科研立项项目(07A064) |
| 摘 要： | T载体能够用于PCR产物的快速克隆且易于操作。以常用的克隆载体pGEM-3zf(+)为出发材料,将其进行改造为通用的T载体。通过一步反向PCR方法在该载体中插入带有Xcm I酶切位点的一段序列,在Xcm I酶的切割下产生两端含有T核苷酸的线型核苷酸序列,最终获得用于克隆PCR产物的T载体。外源基因xdh克隆试验表明该载体构建成功,此载体完全可以用于PCR产物的基因克隆试验,为相关载体的改造提供了思路。 |
| 关 键 词： | T载体 一步反向PCR Xcm I |
Simple Construction Method of a New T-vector and Its Application |
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| Qi Xianghui,Chen Hui,Shen Qi,He Chenxi,Liu Xue,Guo Qi,Chen Huayou,Xu Hong. Simple Construction Method of a New T-vector and Its Application[J]. Biotechnology Bulletin, 2011, 0(3) | |
| Authors: | Qi Xianghui Chen Hui Shen Qi He Chenxi Liu Xue Guo Qi Chen Huayou Xu Hong |
| Affiliation: | Qi Xianghui1,2 Chen Hui1 Shen Qi1 He Chenxi1 Liu Xue1 Guo Qi1 Chen Huayou1 Xu Hong2(1 School of Food and Biological Engineering,Jiangsu University,Zhenjiang 212013,2State Key Laboratory of Materials-Oriented Chemical Engineering,College of Food Science and Light Industry,Nanjing University of Technology,Nanjing 210009) |
| Abstract: | As T-vector can be operated easily,it has been widely used in the rapid cloning of PCR products.pGEM-3zf(+)as a commonly cloning vector,was reconstructed into a common T-vector in this experiment.First of all,the sequence of Xcm I was inserted into pGEM-3zf(+)vector by one-step-reverse PCR,then,the linear nucleotide sequence of pGEM-3zf(+)with T nucleotide at both ends was generated through Xcm I enzyme cutting,eventually,the cloning T-vector used for PCR product was obtained.Cloning experiments of xdh gene... |
| Keywords: | T vector One-step-reverse PCR Xcm I |
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