| 含小鼠PPRE反应元件报告质粒的转化扩增 |
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| 引用本文： | 章涛,张潜,方宁,陈代雄,周岐新.含小鼠PPRE反应元件报告质粒的转化扩增[J].遵义医学院学报,2008,31(1):1-3. |
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| 作者姓名： | 章涛 张潜 方宁 陈代雄 周岐新 |
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| 作者单位： | 1. 遵义医学院附院,贵州省细胞工程重点实验室,贵州,遵义,563003
2. 重庆医科大学,药理学教研室,重庆,400016 |
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| 摘 要： | 目的对含小鼠过氧化物酶体增殖物反应元件（PPRE）报告质粒p4×PPRE-luc进行体外转化和扩增，为基于PPAR为靶标的药物筛选分子平台的建立提供高纯度、高丰度的报告载体。方法取p4×PPRE-luc质粒转化DH5α感受态菌，再接种到含氨苄青霉素的LB琼脂培养板进行抗性筛选培养，挑取阳性克隆菌落进行培养后抽提质粒，紫外分光光度计测定提取质粒的纯度和浓度，并取样进行琼脂糖凝胶电泳验证。结果经转化后获得了抗氨苄青霉素的阳性克隆菌落，挑取阳性克隆并经培养后提取的p4×PPRE—luc质粒其A260/A280=1．84，浓度为750ng／μl。结论成功转化并扩增了p4×PPRE—luc质粒，为基于PPAR为靶标的药物筛选分子平台的建立提供了高纯度、高浓度的报告质粒。
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| 关 键 词： | 过氧化物酶体增殖物反应元件 报告质粒 质粒转化 |
| 文章编号： | 1000-2715(2008)01-0001-02 |
| 修稿时间： | 2007年11月12 |
Transformation and amplification of p4×PPRE-luc reporter vector carrying mouse peroxisome proliferator responsive element |
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| ZHANG Tao,ZHANG Qian,FANG Ning,CHEN Dai-xiong,ZHOU Qi-xin.Transformation and amplification of p4×PPRE-luc reporter vector carrying mouse peroxisome proliferator responsive element[J].Acta Academiae Medicine Zunyi,2008,31(1):1-3. |
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| Authors: | ZHANG Tao ZHANG Qian FANG Ning CHEN Dai-xiong ZHOU Qi-xin |
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| Institution: | ZHANG Tao1,ZHANG Qian1,FANG Ning1,CHEN Dai-xiong1,ZHOU Qi-xin2 (1 Key Laboratory of Cell Engineering of Guizhou Province,The Affiliated Hospital of Zunyi Medical College,Zunyi 563003,China,2 Department of Pharmacology,Chongqing Medical University,Chongqing 400016,China) |
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| Abstract: | Objective To transform and amplify p4×PPRE-luc reporter plasmid carrying mouse pemxisome proliferator responsive element （PPRE） in order to provide a high quality of reporter vector for establishment of drugs screening molecular platform based on mouse PPAR （mPPAR）. Methods p4×PPRE-luc was transformed into DH5α competent cell and cultured in LB agar plate containing ampicillin. Positive clones were selected and cultured in LB liquid medium before plasmid isolating. The purity and concentration of isolated p4×PPRE-luc plasmid from clone culture were analyzed by ultraviolet spectrophotometer. Results The purity and concentration of isolated p4×PPRE-luc plasmid were 1.84（A260/A280） and 750 ng/μ1, respectively. Conclusions Successfully transforming and acquiring high purity and concentration of p4× PPRE-luc reporter plasmid, which provide a good basis for PPAR associated drugs screening. |
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| Keywords: | Peroxisome proliferators responsive element reporter vector plasmid transformation |
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