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目的克隆刺五加GAPDH基因的DNA及启动子序列,并进行生物信息学分析。方法以刺五加的基因组DNA为模板,采用PCR和TAIL-PCR技术克隆GAPDH基因的全长DNA序列及5’端上游启动子序列,并对其进行生物信息学分析。结果克隆到长4 103 bp的刺五加GAPDH基因全长DNA及启动子序列。该基因共包含12个外显子和11个内含子,其剪切均符合GT-AG原则。刺五加GAPDH的启动子片段长1 304 bp,转录起始位点A位于起始密码子ATG上游61 bp处。启动子除含有TATA-box、CAAT-box等基本元件外,还有诸多与激素应答、光响应和胁迫信号等有关的顺式作用调控元件。结论首次克隆到刺五加GAPDH基因的全长DNA及启动子序列,为深入研究GAPDH基因结构特征与功能奠定基础。
Objective To clone the DNA and promoter sequences of GAPDH gene from Acanthopanax senticosus and analyze its bioinformatics. Methods The genomic DNA of Acanthopanax senticosus was used as a template to clone the full-length DNA sequence of GAPDH gene and its upstream 5 ’end promoter sequence by PCR and TAIL-PCR, and its bioinformatics analysis was performed. Results The full-length DNA and promoter sequence of GAPDH gene of Acanthopanax senticosus was cloned. The gene contains a total of 12 exons and 11 introns, the cut is in line with the GT-AG principle. The promoter fragment of GAPDH of Acanthopanax senticosus is 1 304 bp in length, and the transcription start site A is located at 61 bp upstream of the start codon ATG. In addition to promoters containing TATA-box, CAAT-box and other basic components, there are many and hormone response, light response and stress signals and other related cis-acting regulatory elements. Conclusion The full-length DNA and promoter sequence of GAPDH gene in Acanthopanax senticosus was cloned for the first time, which laid the foundation for further study on the structural characteristics and function of GAPDH gene.