| 可用于植物转化的RNAi载体构建方法 |
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| 引用本文： | 娄玲玲,郑唐春,曲冠证,李开隆.可用于植物转化的RNAi载体构建方法[J].安徽农业大学学报,2012,39(1):120-123. |
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| 作者姓名： | 娄玲玲 郑唐春 曲冠证 李开隆 |
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| 作者单位： | 东北林业大学林木遗传育种国家重点实验室,哈尔滨,150040;东北林业大学林木遗传育种国家重点实验室,哈尔滨,150040;东北林业大学林木遗传育种国家重点实验室,哈尔滨,150040;东北林业大学林木遗传育种国家重点实验室,哈尔滨,150040 |
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| 基金项目： | 国家自然科学基金(30972389)资助 |
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| 摘 要： | 利用PCR方法扩增杨树Poptr;CYCD1;1基因的一段内含子序列,并克隆到pROKⅡ载体中,以构建可用于植物特定基因表达干扰的RNAi载体。为了验证载体的效果,构建pCAMBIA-1303载体转化烟草,经检测,转基因烟草能过量表达外源GUS与GFP融合蛋白。把GUS及GFP片段序列连接于RNAi载体,并导入pCAMBIA-1303载体转化烟草。试验结果表明,GUS及GFP干扰序列的导入均能大大降低外源GUS与GFP融合基因的表达。结果说明构建的载体能有效干扰特定基因的表达。
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| 关 键 词： | RNAi 转基因 烟草 |
| 收稿时间： | 7/4/2011 12:00:00 AM |
Re-construct of vector for gene silencing in plants |
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| LOU Ling-ling,ZHENG Tang-chun,QU Guan-zheng and LI Kai-long.Re-construct of vector for gene silencing in plants[J].Journal of Anhui Agricultural University,2012,39(1):120-123. |
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| Authors: | LOU Ling-ling ZHENG Tang-chun QU Guan-zheng and LI Kai-long |
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| Institution: | (State Key Laboratory of Forest Genetics and Tree Breeding,Northeast Forestry University,Harbin 150040) |
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| Abstract: | An intron sequence of Poptr;CYCD1;1 from Populus trichocarpa was ligated into binary vector,pROKⅡ,to obtain the construct for gene silencing in plants.The resulting pRNAi was used to silence the exoge-nous fusion gene,GUS:GFP,in transgenic tobacco transformed with pCAMBIA-1303 vector.The specific se-quence from GUS and GFP genes were cloned into pRNAi,respectively,and both of them reduced the expression of GUS:GFP gene.The results implied that the present pRNAi could be used to silence some genes of interest. |
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| Keywords: | RNAi transgenic plant tobacco |
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