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[摘要]
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。
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[Abstract]
Objective To clone and analyze the full length DNA and promoter sequence of GAPDH gene from Eleutherococcus senticosus. Methods PCR and TAIL-PCR techniques were used to clone the full length DNA sequence and promoter sequence of GAPDH gene of E. senticosus, then these two sequences were analyzed by bioinformatics methods. Results Length of 4 103 bp of E. senticosus GAPDH gene DNA and promoter sequence was cloned. Gene structure analysis showed that it contained 12 exons and 11 introns, and the splicing principles of its exon and intron were consistent with GT-AG; The promoter sequence length was 1 304 bp, and the transcription start site located 61 bp upstream of the initiation codon ATG; The promoter elements such as TATA-box, CAAT-box, as well as many cis-regulatory elements were related to hormone signal response, light response and stress signals. Conclusion The full length DNA and promoter sequence of GAPDH gene in E. senticosus is successfully cloned and reported for the first time, and it provides a stable foundation for further study of GAPDH gene structure and function of E. senticosus.
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[基金项目]
国家自然科学基金资助项目(31570683);河北省教育厅资助科研项目(QN2014102);华北理工大学培育基金(SP201508);华北理工大学大学生创新创业训练计划(X2015171)