| 基于颠茄发根的外源基因表达系统的建立 |
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| 引用本文： | 杨春贤,陈敏,廖志华,谌容,阳义健,张磊.基于颠茄发根的外源基因表达系统的建立[J].园艺学报,2006,33(5):1103-1105. |
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| 作者姓名： | 杨春贤 陈敏 廖志华 谌容 阳义健 张磊 |
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| 作者单位： | (西南大学生命科学学院, 三峡库区生态环境教育部重点实验室, 天然产物与代谢工程实验室, 重庆400715; 第二军医大学药学院生药学教研室, 上海200433) |
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| 基金项目： | 国家自然科学基金资助项目(30500303)，重庆市自然科学基金项目(CSTS2006BA1001)，西南大学博士基金资助项目(SWNUB200411) |
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| 摘 要： | 用携带植物高效表达载体pCAMBIA1304 + “解除武装”的重组C58C1工程菌转化颠茄无菌苗真叶, 发根诱导率达100%。PCR检测证实发根型质粒pRiA4和表达型质粒pCAMBIA1304 +均能整合到颠茄发根基因组内, 共转化率达30%。建立了基于颠茄发根的高效外源基因表达系统, 为将托品烷类生物碱东莨菪碱的生物合成关键酶基因导入颠茄, 实现其次生代谢工程及开展分子育种奠定了基础。
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| 关 键 词： | 颠茄 托品烷类生物碱 发根 外源基因 代谢工程 |
| 文章编号： | 0513-353X（2006）05-1103-03 |
| 收稿时间： | 2005-11-07 |
| 修稿时间： | 2005-11-072006-02-28 |
Establishment of the Exogenous Gene Expression System Based on Atropa belladonna Hairy Roots |
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| Yang Chunxian,Chen Min,Liao Zhihua,Chen Rong,Yang Yijian,Zhang Lei.Establishment of the Exogenous Gene Expression System Based on Atropa belladonna Hairy Roots[J].Acta Horticulturae Sinica,2006,33(5):1103-1105. |
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| Authors: | Yang Chunxian Chen Min Liao Zhihua Chen Rong Yang Yijian Zhang Lei |
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| Institution: | (Laboratory of Natural Products and Metabolic Engineering, Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, School of Life Sciences, Southwest University, Chongqing 400715, China; Department of Pharm acognosy Science, School of Pharmacy, Second Military Medical University, Shanghai 200433, China) |
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| Abstract: | The disarmed Agrobacterium tumefaciens strain C58C1 harboring the plasmid pRiA4 was engineered by introducing the plant expressing vector pCAMBIA1304~ . The bacteria-free well-growing true leaves of A. belladonna were infected with the engineered recombined C58C1. The hairy roots were induced from the wounded sites of the leaves at the frequency of 100%. Then, genomic PCR was applied to detect rolB, rolC, and antihygromycin gene in different monoclonal hairy root lines. All the three genes were simultaneously detected in 30% hairy root clones. The results demonstrated that exogenous genes could be expressed in hairy roots of A. belladonna at a relatively high frequency by the method in the present studies. These were fundamental works for tropane alkaloids metabolic engineering in hairy roots and molecular breeding of A. belladonna. |
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| Keywords: | Atropa belladonna Tropane alkaloids Hairy root Exogenous gene Metabolic engineering Regeneration |
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