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颠茄H6H基因的克隆及高效植物表达载体的构建 被引量:1

Cloning of H6H Gene from Atropa belladonna and Construction of the Efficient Plant Expression Vector
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摘要 [目的]克隆颠茄(Atropa belladonna)H6H基因并构建高效植物表达载体。[方法]采用RT-PCR方法从颠茄中克隆莨菪碱-6β-羟化酶和1,4-丁二胺-氮-甲基转移酶基因编码区,插入经改造后获得双元三价植物高效表达载体p2301-gus,构建植物表达载体p2301-H6H,并将该表达载体导入根癌农杆菌LBA4404和发根农杆菌C58C1。[结果]获得了可直接用于遗传改良的颠茄工程菌p2301-H6H-LBA4404和p2301-H6H-C58C1。[结论]为利用植物基因工程技术提高颠茄中生物碱含量奠定了基础。 [ Objective ] The aim was to clone H6H gene from Atropa belladonna and construct an effective plant expression vector. [ Method J The coding sequence of H6H (Hyoscyamine 613-hydroxylase) was cloned from Atropa belladonna with RT-PCR. Then, the sequence was sub- cloned into the reconstructed plant binary expression vector p2301 to construct the recombinant vector p2301-H6H, which was then introduced into Agrobacterium tumefaciens strain LBA4404 and Agrobacterium rhizogenes strain C58C1, respectively. [ Result] The engineering bacteria p2301-//6H-LBA4404 and p2301-H6H-C58C1 which could be directly used in genetic improvement were obtained. [ Conclusion] The present research provided basis for the increasing of alkaloid content of Atropa belladonna by plant genetic engineering technology.
出处 《安徽农业科学》 CAS 北大核心 2011年第17期10194-10196,共3页 Journal of Anhui Agricultural Sciences
基金 国家"863"计划项目(2010AA100503) 重庆市自然科学基金项目(2009-BB5309) 教育部中央高校基本业务费(XDJK2010C087) 重庆市高等学校优秀人才资助计划项目
关键词 颠茄 莨菪碱-6β-羟化酶 高效植物表达载体 Atropa belladonna Hyoscyamine 6β-hydroxylase Plant expression vector
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