甘蓝Ogura细胞质雄性不育相关基因BoMF1启动子的克隆及功能分析  

Cloning and Functional Analysis of OguCMS-related Gene BoMF1 Promoter in Brassica oleracea

作　　者：郭盈盈[1,2] 颉建明[1] 简元才[2] 郁继华[1] 康俊根[2] 

机构地区：[1]甘肃农业大学农学院,兰州730070 [2]北京市农林科学院蔬菜研究中心,农业部华北地区园艺作物生物学与种质创制重点实验室,北京100097

出　　处：《园艺学报》2013年第5期887-895,共9页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31171958);北京市自然科学基金项目(6102012);‘十二五’国家科技支撑计划项目(2012BAD02B01);‘863’项目(2012AA1001);北京市科委重大科技项目(D111100001311002;D121100003412001)

摘　　要：通过TAIL-PCR染色体步移技术,从甘蓝(Brassica oleracea L.var.capitata L.)基因组中克隆到胞质雄性不育(OguCMS)相关基因BoMF1翻译起始位点上游521bp的启动子序列。软件分析预测表明,该启动子序列中存在多个顺式作用元件,包括TATA-box、CAAT-box、MYB结合位点、植物激素响应单元等。为了研究该启动子的表达特性,亚克隆了BoMF1转录起始位点上游521bp序列,将其置换pBI121中的CaMV35S启动子,驱动其下游的GUS基因,构建植物表达载体pBI121-BoMF1P,以pBI121空载体作为阳性对照,通过农杆菌(LBA4404)介导法转入拟南芥。结果表明,甘蓝BoMF1启动子序列能驱动GUS基因在拟南芥花药发育晚期的花药和花粉中特异表达,表达具有组织特异性。The regulative sequence （521 bp） of OguCMS-related gene BoMF1 promoter from Brassica oleracea was cloned by genomic walking （TAIL-PCR） . In silico analysis showed that this sequence contained several acting elements, including TATA-box and CAAT-box, MYB binding sites, phytohormone responsive elements and so on. In order to study the promoter function, a 521 bp promoter sequence of BoMF1 was inserted upstream of the GUS reporter gene replacing the CaMV35S promoter of pBI121. The plant expression vector pBI121-BoMF1P and pBI121 were transformed into Arabidopsis thaliana with the Agrobacterium tumefaciens strain LBA4404. It was suggested that pBI121-BoMF1P could drive the GUS gene exclusively express in anther and pollen ofArabidopsis thaliana.

关 键 词：甘蓝 OGUCMS BoMF1启动子 GUS 调控元件 

分 类 号：S635.1[农业科学—蔬菜学]