荔枝多酚氧化酶基因启动子克隆与功能分析  

Cloning and function analysis of the promoter of PPO gene from litchi(Litchi chinensis Sonn.)

作　　者：王树军[1,2] 刘保华[2] 孙进华[2] 肖茜[2] 李焕苓[2] 王家保[2] 

机构地区：[1]海南大学农学院,海口570228 [2]中国热带农业科学院环境与植物保护研究所,海口571101

出　　处：《果树学报》2015年第3期427-433,524,共8页Journal of Fruit Science

基　　金：国家自然科学基金(30960233);国家现代农业产业技术体系(CARS-33-02);中央级公益性科研院所基本科研业务专项资金(中国热带农业科学院热带作物品种资源研究所)(1630032012030)

摘　　要：【目的】获得多酚氧化酶(PPO)基因启动子序列并初步分析其功能,为进一步研究PPO基因表达规律及应用PPO基因启动子提供基础。【方法】通过TAIL-PCR和接头PCR方法相结合获得荔枝PPO基因启动子序列并进行生物信息学分析,序列缺失结合瞬时表达法分析核心启动子调控元件。【结果】从‘妃子笑’荔枝基因组中克隆获得了1048bp的荔枝PPO基因启动子序列,含有多种顺式作用元件。不同长度的启动子序列均能驱动GUS基因在‘妃子笑’、‘无核’和‘紫娘喜’荔枝叶片和果皮中表达,但都不能驱动GUS基因在‘妃子笑’和‘紫娘喜’荔枝种子中表达。【结论】获得了荔枝PPO基因启动子序列并获得了其调控基因表达的部分规律。【Objective】The aim of this paper is to clone the promoter sequence of PPO gene and analyzeits function to provide basis for further study on expression of PPO gene and utilization of its promoter.【Methods】The promoter sequence of the PPO gene from litchi was cloned by the methods of TAIL-PCRand adaptor PCR. The characteristics of the promoter sequence were analyzed with bioinformatics soft-ware. The core promoter regulatory elements were verified by constructing and expressing transient expres-sion vectors containing the full or partial promoter sequence.【Results】The 1048 bp promoter sequence ofthe PPO gene was cloned from‘Feizixiao'litchi. It contained a variety of cis-acting elements. The ex-pression of GUS could be mediated by different length of promoter sequences in leaves and peels of threelitchi cultivars,i.e.,‘Feizixiao',‘Ziniangxi'and‘Wuhe'. However,all the promoter sequences couldnot drive the expression of GUS in seeds of‘Ziniangxi'and‘Feizixiao'.【Conclusion】The promoter of PPO gene has been obtained and some of its rules in regulating gene expression has been elucidated.

关 键 词：荔枝 PPO 启动子 表达分析 

分 类 号：S667.1[农业科学—果树学]