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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 promoter sequence of 521bp upstream of the translation initiation site of BoMF1 gene was cloned from the Brassica oleracea L.var.capitata L. genome by TAIL-PCR. Software analysis and prediction showed that there are multiple cis-acting elements in the promoter sequence, including TATA-box, CAAT-box, MYB binding site, plant hormone response unit and so on. In order to study the expression characteristics of this promoter, a 521bp upstream of the transcriptional start site of BoMF1 was subcloned to replace the CaMV35S promoter in pBI121 and drive the downstream GUS gene. The plant expression vector pBI121-BoMF1P was constructed and pBI121 null The vector was used as a positive control and transformed into Arabidopsis by Agrobacterium (LBA4404) mediated method. The results showed that the promoter sequence of BoMF1 in Brassica oleracea could drive the expression of GUS gene in the anther and pollen of Arabidopsis late anther development.