刚毛柽柳水通道蛋白ThPIP基因启动子的克隆及其活性分析

Cloning and GUS Activity Analysis of the Promoter of ThPIP Gene from Tamarix hispida

[目的]研究刚毛柽柳水通道蛋白ThPIP基因启动子的克隆及其活性分析。[方法]采用染色体步移技术,克隆到刚毛柽柳ThPIP基因起始密码子上游1 241 bp启动子序列,并通过PLACE和PlantCARE预测启动子序列中所包含的主要顺式作用元件。将该启动子替换pCAMBIA1301上的35S启动子,构建融合表达基因proThPIP∷GUS,通过基因枪瞬时转化烟草,并进行GUS组织化学染色。[结果]该启动子具有活性,能够驱动GUS基因的表达。[结论]为进一步鉴定该启动子中的顺式作用元件及相互作用的调控因子奠定基础,从而为揭示刚毛柽柳ThPIP基因的分子调控机制提供依据。

[Objective] The aim was to study cloning and GUS activity analysis of the promoter of ThPIP gene trom Tamarix hispida.[Method] A 1241 bp promoter sequence of upstream of initial codon of ThPIP gene was obtained from Tamarix hispida by using the genome walking.Sequence analysis of the promoter region revealed several cis-activity elements in the promoter using the PLACE and PlantCARE.Further,the 35S promoter in pCAMBIA1301 was replaced with the ThPIP promoter to drive the β-glucuronidase （GUS） gene,the proThPIP ∷ GUS recombinant construct was generated,which was transient expressed in tabacco leaves by the particle bombardment method.[Result] The histochemical GUS staining showed that the ThPIP promoter had activity and could drive the expression of GUS gene in tabacco leaves.[Conclusion] The study laid the foundation for further identifing the components and regulatory factors of interactions cis,in order to provide the basis for revealing regulatory mechanism of the ThPIP gene.