| 破色期番茄果实均一化cDNA沉默文库的构建和功能基因筛选模型的初步建立 |
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| 引用本文： | 王晓光,马远征,王晓辉,李玲,罗云波,朱本忠.破色期番茄果实均一化cDNA沉默文库的构建和功能基因筛选模型的初步建立[J].生物化学与生物物理进展,2009,36(8):1079-1083. |
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| 作者姓名： | 王晓光 马远征 王晓辉 李玲 罗云波 朱本忠 |
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| 作者单位： | 中国农业大学食品科学与营养工程学院食品生物技术实验室,北京,100083 |
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| 基金项目： | 国家自然科学基金资助项目(30600421, 30871741) |
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| 摘 要： | 番茄果实的成熟是由多基因精细调控的一个过程．利用破色期番茄果实，根据复性动力学原理在mRNA水平进行均一化操作使高丰度和低丰度的mRNA丰度接近．然后把均一化之后mRNA反转录得到cDNA，再与基因沉默载体pTRV重组，最后把构建好的载体通过电转化的方法转入到GV3101农杆菌中，从而建立起破色期番茄果实均一化cDNA沉默文库．通过番茄果实中病毒诱导基因沉默技术，对cDNA沉默文库进行初步筛选，从而确定功能基因筛选模型．在模型建立阶段，以番茄红素合成途径相关的PDS基因作为内标基因，在100个混合农杆菌样中，成功筛选到了PDS基因．
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| 关 键 词： | cDNA文库，均一化，功能基因，VIGS |
| 收稿时间： | 2008/12/6 0:00:00 |
| 修稿时间： | 3/9/2009 12:00:00 AM |
Construction of a normalized cDNA silencing library of tomato fruit and model establishment of screening specific functions of genes |
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| WANG Xiao-Guang,MA Yuan-Zheng,WANG Xiao-Hui,LI Ling,LUO Yun-Bo and ZHU Ben-Zhong.Construction of a normalized cDNA silencing library of tomato fruit and model establishment of screening specific functions of genes[J].Progress In Biochemistry and Biophysics,2009,36(8):1079-1083. |
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| Authors: | WANG Xiao-Guang MA Yuan-Zheng WANG Xiao-Hui LI Ling LUO Yun-Bo and ZHU Ben-Zhong |
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| Institution: | Food Biotechnology Laboratory, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China;Food Biotechnology Laboratory, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China;Food Biotechnology Laboratory, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China;Food Biotechnology Laboratory, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China;Food Biotechnology Laboratory, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China;Food Biotechnology Laboratory, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China |
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| Abstract: | |
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| Keywords: | cDNA library normalization functional genes VIGS |
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