| 马铃薯块茎糖基转移酶基因的克隆及其RNAi载体的构建 |
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| 引用本文： | 王旺田,张金文,王蒂,张俊莲,司怀军,陶士珩.马铃薯块茎糖基转移酶基因的克隆及其RNAi载体的构建[J].作物学报,2011,37(11):1926-1934. |
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| 作者姓名： | 王旺田 张金文 王蒂 张俊莲 司怀军 陶士珩 |
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| 作者单位： | 1 甘肃农业大学农学院, 甘肃兰州 730070; 2 甘肃省作物遗传改良与种质创新重点实验室, 甘肃兰州 730070; 3 西北农林科技大学生命科学学院, 陕西杨凌 712100 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)项目(2010CB134404); 甘肃省自然科学基金项目(0710RJZA088); 教育部博士生国内访学项目(Z107-020901)资助 |
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| 摘 要： | 为探讨鼠李糖基转移酶(rhamnosyltransferase, sgt3)基因与类固醇糖苷生物碱(steroidal glycoalkaloid, SGAs)合成关系, 揭示sgt3基因在SGAs合成中的作用, 本研究根据GenBanK No：DQ266437保守区设计特异引物, 通过RT-PCR技术获得马铃薯块茎sgt3相似基因；采用生物信息学相关软件分析, 预测sgt3相似基因cDNA序列编码的蛋白质结构和功能, 构建基于sgt3基因的植物干扰表达载体。结果表明, 克隆的sgt3相似基因与报道的sgt3基因序列相似性达到99.54%, 其ORF长1 500 bp, 编码505个氨基酸残基, 具有UDPG糖基转移酶保守结构域及许多重要功能位点；三级结构预测表明, 该氨基酸同糖基转移酶单体结构模型相似, 为糖基转移酶家族成员, 表明可能具有合成类固醇糖苷生物碱功能, 基因序列已注册到GenBank, 序列登录号为HM188447。以此为靶序列, 构建由Actin启动子和CIPP启动子驱动的与sgt1和sgt2基因相似度高的含有sgt3基因序列的干扰表达载体, 将为糖苷生物碱的合成、代谢的进一步研究及低糖苷生物碱转基因马铃薯品种的培育奠定基础。
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| 关 键 词： | 马铃薯 鼠李糖基转移酶 基因克隆 序列分析 RNAi载体 |
| 收稿时间： | 2011-04-27 |
Cloning of Rhamnosyl Transferase Gene and Construction of Its RNAi Vector in Potato |
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| WANG Wang-Tian,ZHANG Jin-Wen,WANG Di,ZHANG Jun-Lian,SI Huai-Jun, TAO Shi-Heng College of Agronomy,Gansu Agricultural University,Lanzhou ,China, Gansu Key Laboratory of Crop Genetic & Germplasm Enhancement, College of Life Sciences,Northwest A&F University,Yangling.Cloning of Rhamnosyl Transferase Gene and Construction of Its RNAi Vector in Potato[J].Acta Agronomica Sinica,2011,37(11):1926-1934. |
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| Authors: | WANG Wang-Tian ZHANG Jin-Wen WANG Di ZHANG Jun-Lian SI Huai-Jun TAO Shi-Heng College of Agronomy Gansu Agricultural University Lanzhou China Gansu Key Laboratory of Crop Genetic & Germplasm Enhancement College of Life Sciences Northwest A&F University Yangling |
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| Institution: | 1.College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China;2.Gansu Key Laboratory of Crop Genetic & Germplasm Enhancement, Lanzhou 730070, China;3.College of Life Sciences, Northwest A&F University, Yanglin 712100, China |
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| Abstract: | Steroidal glycoalkaloids (SGAs) are potentially harmful metabolites found in potatoes and other Solanaceous plants. SGAs accumulation affects food quality and safety, we hope to use molecular biology methods to reduce SGAs content and assist breeding efforts to ensure food safety, develop new and improved varieties of potatoes. To investigate the relationship between the gene expression of the rhamnosyltransferase (sgt3) and the biosynthesis of steroidal glycoalkaloid (SGAs), and reveal the role of sgt3 gen... |
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| Keywords: | Potato sgt3 gene Gene cloning Sequence analysis RNA interference vector |
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