Construction and confirmation of tetracycline-dependent regulatory system in one vector

ZHOU Zhi; et al

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Construction and confirmation of tetracycline-dependent regulatory system in one vector

更新时间：2024-01-17

- Construction and confirmation of tetracycline-dependent regulatory system in one vector
- Journal of Chongqing Medical University Issue 1, Pages: 23-25(2006)
- 作者机构：
  
  1. 重庆医科大学附属第二医院肝病治疗研究中心
  2. 重庆医科大学附属第二医院肝病治疗研究中心,重庆,400010
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2006，
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[1]周智,倪艳,雷宇,王蜀强,陈压西,任红.四环素双表达调控表达载体的构建、鉴定[J].重庆医科大学学报,2006(01):23-25+94.

ZHOU Zhi, et al. Construction and confirmation of tetracycline-dependent regulatory system in one vector. [J]. Journal of Chongqing Medical University (1):23-25(2006)
[1]周智,倪艳,雷宇,王蜀强,陈压西,任红.四环素双表达调控表达载体的构建、鉴定[J].重庆医科大学学报,2006(01):23-25+94. DOI：

ZHOU Zhi, et al. Construction and confirmation of tetracycline-dependent regulatory system in one vector. [J]. Journal of Chongqing Medical University (1):23-25(2006) DOI：

摘要

目的:构建在同一载体中可用四环素调控表达目的基因的载体。方法:以pcDNA4TM/TO为模板

用PCR法扩增含四环素调控表达序列的片段TO（位于NdeI和SacII之间）

插入pUC118载体中测序

选择测序正确的序列取代pVITRO3载体中相应的片段

构建成pVITRO3-TO。用相同的方法以pcDNA6/TR○c为模板扩增四环素阻遏蛋白基因（TR）

经测序证实后插入pVITRO3-TO载体的一个多克隆位点。结果:用PCR成功扩增出四环素调控表达序列片段和四环素阻遏蛋白基因片段

亚克隆后各选2个克隆测序

均找到了序列正确的克隆

经内切酶双切后插入pVITRO3中

完成了目的载体的构建。结论:初步构建了四环素双表达调控表达载体（pVITRO3-TO-TR）。

Abstract

Objective: To construct tetracycline-dependent regulatory system in one vector

which contains two multiple cloning sites(MCS) for the convenient cloning of two target genes.Methods: The tetracycline(Tet)-dependent regulatory fragment(TO)(between NdeI and SacII sites) was amplified from pcDNA4TM/TO vector with PCR

which was inserted into pUC118 vector and sequenced.The counterpart fragment in pVITRO3 was replaced with correct Tet-dependent regulatory fragment

namely pVITRO3-TO vector.The Tet repressor gene(TR) encoding the Tet repressor from pcDNA6/TR vector was incorporating into the backbone of the pVITRO3-TO.Result: The tetracycline(Tet)-dependent regulatory fragment and the Tet repressor gene were amplified successfully with PCR from its template vectors and were subcloned.The clones with correct sequence were selected by sequencing the picked 2 clones.The target vector was constructed successfully by incorporating TO and TR into the backbone of pVITRO3 vector.Conclusion: Tetracycline-dependent regulatory double-expressing system in one vector was constructed initially.

关键词

调控表达载体构建四环素

Keywords

Regulatory expressing vectorconstructiontetracycline

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