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[目的]将牻牛儿基牻牛儿基焦磷酸合成酶(geranylgeranyl pyrophosphate synthase,GGPS)基因转化西瓜。[方法]利用RT-PCR技术克隆西瓜果实GGPS基因CDS区;构建由果实特异性启动子AGPL1调控GGPS基因、以nptⅡ为筛选标记基因的植物表达载体,并转入农杆菌EHA105;通过农杆菌介导法对西瓜品种“红一号”进行遗传转化。[结果]经过卡那霉素筛选和PCR初步鉴定,获得4株转基因阳性植株。[结论]构建了GGPS基因果实特异性表达载体,并将其转入西瓜品种“红一号”中,转化效率为0.13%。
[Objective] The research aimed to transform geranylgeranyl pyrophosphate synthase (GGPS) gene into watermelon. [Method] The CDS region of GGPS gene in watermelon fruit was cloned by RT-PCR. The GGPS gene was regulated by fruit-specific promoter AGPL1, and the plant expression vector with nptⅡ as selectable marker gene was transformed into Agrobacterium tumefaciens EHA105. Guide method of watermelon varieties “Red One ” for genetic transformation. [Result] After kanamycin screening and preliminary identification by PCR, 4 transgenic plants were obtained. [Conclusion] The fruit-specific expression vector of GGPS gene was constructed and transferred into watermelon cultivar “Red No.1” with the transformation efficiency of 0.13%.