| 细菌-酵母穿梭表达质粒pGBKT7-LMP1 的构建与鉴定 |
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| 引用本文： | 刘鹏,张清秀,朱振宇,庾蕾,马涧泉.细菌-酵母穿梭表达质粒pGBKT7-LMP1 的构建与鉴定[J].中山大学学报(医学科学版),2001,22(5):348-351. |
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| 作者姓名： | 刘鹏 张清秀 朱振宇 庾蕾 马涧泉 |
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| 作者单位： | 1. 中山医科大学生化教研室,
2. 中山医科大学法医系, |
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| 基金项目： | 广东省自然科学基金资助项目(994004) |
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| 摘 要： | 目的]构建能在酵母中表达EBV潜伏膜蛋白1(LMPl)的穿梭表达质粒pGBKT7-LMP1.方法]RT-PCR方法扩增EB病毒LMP1全外显子片段,利用DNA重组技术将其定向插人细菌-酵母穿梭质粒pGBKT7的T7启动子下游.结果]限制性内切酶酶切和DNA测序分析证实RT-PCR获得的LMP1 cDNA片段与GenBank中的数据完全吻合;LMP1准确克隆人pGBKT7的多克隆位点,未改变读码框架.结论]成功构建了穿梭质粒pGBKT7-LMP1.
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| 关 键 词： | 疱疹病毒4型 人 潜伏膜蛋白1 聚合酶链反应/方法 质粒pGBKT7 |
| 文章编号： | 1000-257X(2001)05-0348-04 |
| 修稿时间： | 2000年12月12 |
Construction and Identification of Bacteria-Yeast Shuttle Plasmid pGBKT7-LMP1 |
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| LIU Peng ,ZHANG Qing xiu ,ZHU Zhen yu ,YU Lei ,MA Jian quan.Construction and Identification of Bacteria-Yeast Shuttle Plasmid pGBKT7-LMP1[J].Journal of Sun Yatsen University(Medical Sciences),2001,22(5):348-351. |
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| Authors: | LIU Peng ZHANG Qing xiu ZHU Zhen yu YU Lei MA Jian quan |
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| Affiliation: | LIU Peng 1,ZHANG Qing xiu 1,ZHU Zhen yu 1,YU Lei 2,MA Jian quan 1 |
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| Abstract: | To construct bacteria yeast shuttle plasmid pGBKT7 LMP1. The fragment including all the exons of LMP1 was amplified by RT PCR and was recombined to the downstream of T7 promoter in the shuttle vector pGBKT7. It has been proved that the sequence of the RT PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The LMP1 cDNA fragment was cloned in the vector pGBKT7 in the right direction, and the open reading fragment of LMP1 was maintained. Conclusion] The pGBKT7 LMP1, a bacteria yeast shuttle plasmid is successfully constructed. |
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| Keywords: | herpesvirus 4 human latent membrane protein 1 polymerase chain reaction/methods plasmid pGBKT7 |
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