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[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(Atropa belladonna)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。
[Objective] The research aimed to clone the H6H gene of Atropa belladonna and construct a high efficient plant expression vector. [Method] The coding region of scopolamine-6β-hydroxylase and 1,4-butanediamine-nitrogen-methyltransferase gene was cloned by RT-PCR from Atropa belladonna, Yuan three-plant efficient expression vector p2301-gus, construct plant expression vector p2301-H6H, and the expression vector was introduced into Agrobacterium tumefaciens LBA4404 and Agrobacterium rhizogenes C58C1. [Result] The strains of Helicoverospora such as p2301-H6H-LBA4404 and p2301-H6H-C58C1 that could be directly used for genetic improvement were obtained.