| 植物亚细胞定位载体卡盒pCEG的构建及验证 |
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| 引用本文： | 朱丹,王希,朱延明,陈超,李勇,柏锡,才华,纪巍.植物亚细胞定位载体卡盒pCEG的构建及验证[J].东北农业大学学报,2011,42(4):83-88. |
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| 作者姓名： | 朱丹 王希 朱延明 陈超 李勇 柏锡 才华 纪巍 |
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| 作者单位： | 东北农业大学生命科学学院,哈尔滨,150030 |
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| 基金项目： | 国家高科技发展计划(863计划)，国家自然科学基金资助项目，黑龙江省科技厅重大攻关项目，黑龙江省教育厅科技项目，黑龙江省教育厅科技项目 |
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| 摘 要： | GFP基因被广泛地应用于植物转基因以及基因功能验证研究,然而构建与GFP融合的植物表达载体,常会因酶切位点难以选择,而使得载体构建过程复杂,周期长.以含GFP的质粒pCAMBIA-1302为基础,消除此质粒本身的多克隆位点(MCS),并在此质粒GFP基因序列前插入原核表达载pET-32b的多克隆位点,构建了植物GFP亚...
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| 关 键 词： | 载体卡盒构建 亚细胞定位 GFP 烟草 |
Construction and application of a plant subcellular localization vector box of pCEG |
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| ZHU Dan,WANG Xi,ZHU Yanming,CHEN Chao,LI Yong,BAI Xi,CAI Hua,JI Wei.Construction and application of a plant subcellular localization vector box of pCEG[J].Journal of Northeast Agricultural University,2011,42(4):83-88. |
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| Authors: | ZHU Dan WANG Xi ZHU Yanming CHEN Chao LI Yong BAI Xi CAI Hua JI Wei |
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| Institution: | (College of Life Sciences,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | GFP gene was widely applied in the transgenic plants and genes function’s proving study.However,the construction of GFP fused plant expression vectors was always complicated and time consuming due to the hard choose of restriction enzyme cutting sites.Use plasmid pCAMBIA-1302 which included GFP gene as a original vector,and eliminated its own multiple clone site(MCS),and then inserted pET-32b’s multiple clone site in front of GFP gene sequences,named the eventual plant GFP subcellular localization vector box pCEG.By Agrobacterium-mediated transformation method,pCEG instantaneous expressed in tobacco leaves,and GFP protein subcellular localization was observed by laser confocal microscope.The result displayed that the GFP proteins expressed in both cytoplasm and nucleus,this means the pCEG box can be used conveniently for plant genes subcellular localization analysis and provide a lot of usable restriction enzyme cutting site for plant genes fusioned with GFP,and the pCEG box is convenient for plant genes function analysis and has great use value. |
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| Keywords: | GFP |
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