| Gateway技术快速构建百合双价抗病毒RNAi表达载体 |
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| 引用本文： | 徐品三,白建芳,刘纪文,李焕改.Gateway技术快速构建百合双价抗病毒RNAi表达载体[J].中国农学通报,2011,27(4):144-147. |
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| 作者姓名： | 徐品三 白建芳 刘纪文 李焕改 |
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| 作者单位： | 大连理工大学生命科学与技术学院,辽宁,大连,116023 |
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| 摘 要： | 为获得适合转化百合的双价RNAi表达载体，以感病百合叶片的总RNA为模板，分别扩增400 bp左右的LSV和LMoV CP基因。通过重叠延伸PCR技术获得长约800 bp的LSV-LMoV融合基因。利用Gateway技术，通过BP反应及LR反应将LSV-LMoV融合基因克隆到双元载体pH7GWIWG2(Ⅱ)，成功构建了含两种病毒基因的RNAi表达载体pH7GWIWG2(Ⅱ)-LSV-LMoV。将构建好的双价表达载体导入农杆菌EHA105，PCR和测序表明所构建的载体及获得的工程菌与预期的完全一致。
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| 关 键 词： | 干旱监测评估 干旱监测评估 内蒙古地区 服务系统 |
| 收稿时间： | 9/3/2010 12:00:00 AM |
| 修稿时间： | 2010/11/2 0:00:00 |
Construction of LSV and LMoV Binary Virus Resistant RNAi Vector Using Gateway Technology |
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| Xu Pinsan,Bai Jianfang,Liu Jiwen,Li Huangai.Construction of LSV and LMoV Binary Virus Resistant RNAi Vector Using Gateway Technology[J].Chinese Agricultural Science Bulletin,2011,27(4):144-147. |
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| Authors: | Xu Pinsan Bai Jianfang Liu Jiwen Li Huangai |
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| Institution: | Xu Pinsan,Bai Jianfang,Liu Jiwen,Li Huangai(School of Life Science and Biotechnology,Dalian University of Technology,Dalian Liaoning 116023) |
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| Abstract: | In order to gain the RNA interference（RNAi） transformation vector of lily,two fragment genes about400 bp of LSV and LMoV were obtained from the total RNA of lily leaves infected.LSV-LMoV fusion gene was constructed by recombinant PCR technique.Using Gateway cloning technology,the RNAi transformation vector pH7GWIWG2（Ⅱ）was constructed by BP and LR recombination reactions.PCR and sequencing analysis confirmed that the pH7GWIWG2（Ⅱ）-LSV-LMoV vector,a binary vector containing two different virus genes,was obtained successfully.Virus susceptible lily was transformed with pH7GWIWG2（Ⅱ）-LSV-LMoV by Agrobacterium tumefaciens-mediated transformation. |
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| Keywords: | lily symptomless virus lily mottle virus RNAi vector recombinant PCR gateway technology |
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