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

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.