白桦BpEIN3基因启动子克隆与表达特性

Cloning and Expression Analysis of EIN3 Promoter in Betula platyphylla×B.pendula

EIN3基因是乙烯信号转导中重要的转录因子,在植物的衰老中发挥重要的作用。为研究EIN3基因在白桦中的表达特性,通过PCR方法克隆了2843 bp白桦BpEIN3基因的启动子区域片段,构建了pBpEIN3::CUS表达载体进行白桦转基因,获得1株pBpEIN3::CUS白桦株系;并利用PLANCE在线软件对BpEIN3基因启动子片段的主要顺式作用元件进行的分析预测;同时分别用ET、IAA、MeJA、SA和GA处理转pBpEIN3::GUS白桦株系,通过实时定量PCR及GUS活性的检测分析该启动子的表达调控模式。结果表明,BpEIN3启动子包含着多种功能的调控顺式元件;且pBpEIN3::GUS对不同激素均有不同程度的响应,其中SA、GA响应最为明显,说明不同激素对BpEIN3基因的表达模式产生了影响。综上表明:所克隆的BpEIN3基因上游2843 bp的DNA片段具有启动子活性,且能驱动报告基因的有效表达,为后续基因功能的研究奠定基础。

Ethylene insensitive 3(EIN3)is an essential transcription factor of the ethylene signal transduction,which plays an important role in plant senescence.To identify the expression characteristics of EIN3 gene in birch,the promoter fragment of BpEIN3 about 2843 bp was cloned by using PCR.The sequence elements of BpEIN3 promoter fragment were analyzed by PLANCE.The pBpEIN3::GUS vector was constructed and transferred into birch,and one transgenic line with GUS activity was obtained.Further,the expression and activity of GUS in transgenic line under the treatment of ET,IAA,MeJA,SA and GA were detected to explore the response patterns.The results showed BpEIN3 promoter contained multiple cis-regulating elements,which led the transgenic line response to different hormones.The perceptions of transgenic line to SA and GA were significant higher than to other hormones.In summary,the 2843 bp DNA fragment has promoter activity,and it can induce the expression of GUS reporter.