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.
出处
《园艺学报》
CAS
CSCD
北大核心
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)