西瓜GGPS基因果实特异性表达载体构建与遗传转化  

Construction of fruit-specific expression vector of GGPS gene and genetic transformation in watermelon(Citrullus lanatus)

作　　者：吕品[1,2] 李朋伟[1] 马双武[3] 赵文恩[2] Pin Lv Pengwei Li Shuangwu Ma Wen＇ en Zhao(College of Pharmacy, Henan university of Chinese Medicine, Zhengzhou 450008, China School of Chemical Engineering and Energy, Zhengzhon University, Zhengzhou 450001, China Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009, China)

机构地区：[1]河南中医药大学药学院,河南郑州450008 [2]郑州大学化工与能源学院,河南郑州450001 [3]中国农业科学研究院郑州果树研究所,河南郑州450009

出　　处：《生物技术》2016年第5期432-437,455,共7页Biotechnology

基　　金：河南省科技发展计划(基础与前沿技术研究)项目("西瓜果实类胡萝卜素合成积累的代谢调控";No.132300410092)

摘　　要：[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 ] Genetic transformation of geranylgeranyl pyrophosphate synthase （GGPS） gene into watermelon. [ Methods] The coding sequence of GGPS gene was cloned from watermelon fruit using RT - RCR. The plant expression vectors harboring GGPS gene, directed by watermelon fruit -specific expression AGPLlpromoter, was constructed and transferred into Agrobacterium tumefacien EHA105. The GGPS gene was introduced into watermelon cuhivar ＂HongYiHao＂ via Agrobacterium - mediated transformation. [ Results] Four transgenie plants with selective marker gene, npt Ⅱ , were confirmed by PCR analysis among the regenerated plants. [ Conclusion]The fruitspecific expression vectors of GGPS gene was successfully constructed and transferred into watermelon cuhivar ＂HongYiHao＂. The transformation efficiency was 0.13%.

关 键 词：西瓜 GGPS基因 AGPL1启动子 农杆菌介导 遗传转化 

分 类 号：Q785[生物学—分子生物学]