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为了给转基因育种提供诱导型和组织特异型启动子,本研究根据陆地棉GhRCA1基因的cDNA序列和雷蒙德氏棉的基因组序列,采用PCR技术对GhRCA1基因的5’端上游转录调控序列扩增,获得棉花GhRCA1基因的2个启动子序列,分别命名为RCA11和RCA12。生物信息学分析表明,这2个序列均具有多个启动子的基本元件TATA-box和CAAT-box,并含有多种光调控相关的顺式作用元件、激素响应元件、逆境胁迫诱导相关的响应元件等。推测GhRCA1基因的2个启动子都受光的诱导,启动子序列上核苷酸和顺式元件的差异可能使GhRCA1基因的表达受到不同程度的调控。本研究通过克隆和分析GhRCA1基因的启动子序列,为进一步研究该启动子的功能以及基因的表达调控机制奠定了基础。
In order to provide inducible and tissue-specific promoters for transgenic breeding, based on the cDNA sequence of GhRCA1 and the genomic sequence of Raymond cotton, the upstream and downstream transcriptional regulatory sequences of GhRCA1 gene were amplified by PCR , Two promoter sequences of cotton GhRCA1 gene were obtained and named RCA11 and RCA12 respectively. Bioinformatics analysis showed that these two sequences all have multiple promoter elements TATA-box and CAAT-box, and contain a variety of cis-acting elements related to light regulation, hormonal response elements, and stress-induced responses Components and so on. It is speculated that the two promoters of GhRCA1 gene are all induced by light. The differences of nucleotide and cis-element of the promoter sequence may regulate the expression of GhRCA1 gene to some extent. In this study, cloning and analysis of the promoter sequence of GhRCA1 gene laid the foundation for further study on the function of the promoter and the regulation of gene expression.