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利用荧光定量PCR技术对蜡梅花发育的不同组织、不同花期和不同胁迫下的水通道蛋白基因CpAQP1的表达量进行分析.结果表明:CpAQP1在盛开期中瓣的表达量明显高于其他组织器官,在高温、高盐、ABA胁迫下表达量变化明显,说明该基因具有组织表达差异性及能够对环境胁迫做出响应.同时利用hi-TAIL PCR方法克隆获得蜡梅水通道蛋白基因CpAQP1上游启动子1082bp,命名为CpAQP1pro(GenBank登录号为:JQ952563).该序列具有典型的基本元件TATA-box、CAAT-box及与胁迫相关的元件和光应答元件.将克隆的启动子CpAQP1pro替换pBI121中的CaMV35s启动子,构建植物表达载体pBI121-CpAQP1pro,通过农杆菌转化烟草,稳定表达结果说明该启动子具有驱动GUS报告基因表达的活性.
The quantitative analysis of CpAQP1 expression in different tissues, different flowering periods and different stresses of C. przewalskii by quantitative real-time PCR showed that the expression of CpAQP1 in the mid-flowering stage was significantly higher than that in other tissues and organs High temperature, high salt, ABA stress expression changes significantly, indicating that the gene has a difference in tissue expression and can respond to environmental stress.At the same time using the method of cloning cloning and expression of chingmei aquaporin CpAQP1 upstream promoter 1082bp , Named CpAQP1pro (GenBank accession number: JQ952563) .This sequence has the typical basic elements of TATA-box, CAAT-box and stress-related elements and photo-responsive elements.The cloned promoter CpAQP1pro replaced the CaMV35s promoter in pBI121 The plant expression vector pBI121-CpAQP1pro was constructed and transformed into tobacco by Agrobacterium tumefaciens. The stable expression results indicated that the promoter has the activity of driving GUS reporter gene expression.