| 东亚三角涡虫DjPreb基因干扰载体的构建及干扰效果鉴定 |
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| 引用本文： | 袁佐清,赵博生. 东亚三角涡虫DjPreb基因干扰载体的构建及干扰效果鉴定[J]. 生物技术通报, 2011, 0(9) |
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| 作者姓名： | 袁佐清 赵博生 |
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| 作者单位： | 山东理工大学生命科学学院,淄博,255049 |
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| 基金项目： | 山东省自然科学基金项目(ZR2009DM029); 山东理工大学博士启动基金项目(4041-410013) |
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| 摘 要： | 以含有东亚三角涡虫DjPreb基因的pcDNA3-DjPreb重组质粒为模板,经PCR扩增目的片段,将其克隆到干扰载体L4440上,构建重组质粒L4440-DjPreb后转化入大肠杆菌HT115感受态细胞中,IPTG诱导表达dsRNA后喂食涡虫.显微观察喂食dsRNA后的涡虫在再生过程中的表型变化,Real-time PCR检测载体对涡虫DjPreb基因的表达抑制效果.试验结果显示DjPreb基因的RNA干扰表达载体构建成功,DjPreb基因RNA干扰后涡虫不能正常再生.Real-time PCR分析饲喂RNA干扰食物后DjPrcb mRNA的表达显著下降,进一步说明DjPreb在涡虫头尾的形成中发挥作用.
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| 关 键 词： | 涡虫 DjPreb RNA干扰 再生 |
Construction of Interference Vector of Planarian Dugesia japonica DjPreb Gene and Preliminary Identification of Its Interference Efficiency |
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| Yuan Zuoqing,Zhao Bosheng. Construction of Interference Vector of Planarian Dugesia japonica DjPreb Gene and Preliminary Identification of Its Interference Efficiency[J]. Biotechnology Bulletin, 2011, 0(9) |
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| Authors: | Yuan Zuoqing Zhao Bosheng |
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| Affiliation: | Yuan Zuoqing Zhao Bosheng (School of Life Sciences,Shandong University of Technology,Zibo 255049) |
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| Abstract: | cDNA fragment for the DjPreb from adult planarians Dugesia japonica cDNA library was amplified from recombinant plasmid pcDNA3-DjPreb using PCR and then cloned into the interference vector L4440.The recombinant plasmid L4440-DjPreb was transferred into E.coli HT115 to make dsRNA induced by IPTG.RNA interference(RNAi)food was fed directly to worms then DjPreb RNAi animals were amputated into three fragments and allowed to each piece regenerate.Animals were observed with a stereomicroscope during the process ... |
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| Keywords: | Planarian DjPreb RNA interference Regeneration |
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