| 一种新的乙型肝炎病毒全基因组克隆技术的建立 |
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| 引用本文： | 李莉,卢建溪,谢冬英,徐启桓,朱明芬,陈伟,李刚. 一种新的乙型肝炎病毒全基因组克隆技术的建立[J]. 热带医学杂志, 2006, 6(8): 868-870 |
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| 作者姓名： | 李莉 卢建溪 谢冬英 徐启桓 朱明芬 陈伟 李刚 |
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| 作者单位： | 中山大学附属第三医院感染科,广州,510630;中山大学附属第三医院中心实验室,广州,510630 |
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| 基金项目： | 广东省教育厅“千百十工程”优秀人才培养基金(No.Q02034)，广东省医学科研基金(No.A2003222)，教育部新世纪优秀人才支持计划(No.NCET-04-0797)。 |
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| 摘 要： | 目的建立一种乙型肝炎病毒(HBV)全基因组扩增及克隆的新方法。方法设计含SalⅠ、NotⅠ酶切位点的引物,用一片段PCR法扩增HBV全基因组,经双酶切后将PCR产物克隆入pBlueScript-(SK-)载体,进行HBV全基因组序列测定。结果用此方法成功获得35例HBV全基因组DNA序列。同时以重组质粒为模板进行敏感性和保真性测定,其敏感性为102个初始模板,核苷酸的人为突变率为1.6bp/kb。结论此方法可用于大规模HBV全基因组扩增和序列分析,为HBV的基础和临床研究提供了一种新方法。
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| 关 键 词： | 肝炎病毒 乙型 基因组 聚合酶链反应 |
| 文章编号： | 1672-3619（2006）08-0868-03 |
| 收稿时间： | 2006-04-09 |
| 修稿时间： | 2006-05-26 |
A New Method for Amplification and Cloning of the Full-Length HBV Genome |
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| LI Li,LU Jian-xi,XIE Dong-ying,XU Qi-huan,ZHU Ming-fen,CHEN Wei,LI Gang. A New Method for Amplification and Cloning of the Full-Length HBV Genome[J]. Journal Of Tropical Medicine, 2006, 6(8): 868-870 |
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| Authors: | LI Li LU Jian-xi XIE Dong-ying XU Qi-huan ZHU Ming-fen CHEN Wei LI Gang |
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| Abstract: | Objective To develop a new method for amplification and cloning of a full-length HBV genome. Method Primers containing Sal I and Not I sites were used in PCR amplification of the HBV genome. The PCR product was cloned into pBlue Script(SK) plasmid and followed by sequencing. Result 35 clones containing the full-length HBV genome were obtained. The sensitivity and fidelity of this cloning method were also analyzed. The mimium unmber of templates was 102 and the mutation rate was 1.6 bp/kb. Conclusion This method can be used in a large scale amplification and sequence analysis of the full-length HBV genome. |
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| Keywords: | hepatitis B virus genome polymerase chain reaction |
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