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作 者:孙玉萍 王杨[1] 门璟煜 董琦 杨颖[1] 于婷乔 吴晓飞 卢存福[1] 陈玉珍[1] SUN Yu-ping;WANG Yang;MEN Jing-yu;DONG Qi;YANG Ying;YU Ting-qiao;WU Xiao-fei;LU Cun-fu;CHEN Yu-zhen(Beijing Advanced Innovation Center for Tree Breeding by Molecular Design/College of Biological Sciences andBiotechnology, Beijing Forestry University/National Engineering L aboratoiy for Tree Breeding, Beijing 100083;School of Life Sciences, Pecking University, Beijing 100871)
机构地区:[1]北京林木分子设计育种高精尖创新中心/北京林业大学生物科学与技术学院/林木育种国家工程中心,北京100083 [2]北京大学生命科学学院,北京100871
出 处:《植物遗传资源学报》2019年第3期769-780,共12页Journal of Plant Genetic Resources
基 金:国家自然科学基金项目(31270737);高等学校学科创新引智计划项目(B13007);长江学者和创新团队发展计划项目(IRT13047);北京市自然科学基金项目(6112016)~~
摘 要:端粒酶是具有逆转录酶活性的维持真核生物中染色体末端DNA完整性的核糖核蛋白复合物,而端粒酶逆转录酶(TERT,Telomerase reverse transcriptase)是端粒酶的核心组分。目前对TERT启动子的研究主要集中在人体的肿瘤细胞方面,而对植物TERT基因启动子的结构及TERT表达调控机理的研究有所缺乏。本研究通过生物信息学技术对拟南芥TERT启动子结构特征进行了系统分析,并构建植物TERT启动子进化树;用半定量PCR检测野生型和AtTERT启动子区突变体拟南芥幼苗在非生物胁迫下的TRET表达模式。结果表明,系统进化分析显示TERT启动子在同一类群植物中有着较高的保守性;TERT基因启动子具有多个与干旱、盐、激素等相关调控元件;突变体AtTERT转录表达量普遍低于野生型;同时突变体对盐、干旱胁迫抗性降低,而对脱落酸处理不敏感。这说明AtTERT启动子含有抗旱、抗盐反应顺式作用元件,而无响应ABA调控机制,验证了对AtTERT基因启动子的顺式作用元件的预测,并初步证明AtTERT启动子对AtTERT的转录表达具有调控作用,对未来探讨该基因的表达调控机制提供了重要参考。Telomerase is a ribonuclear protein complex with reverse transcriptase activity in eukaryotic cells,with significance on maintaining DNA integrity at the ends of eukaryotic linear chromosomes.The telomerase reverse transcriptase(TERT)is the key subunit of telomerase.Currently,researches on characterization of the TERT promoter is mainly focused on tumor cells,and studies of plant TERT promoter remain unclear to date.In this study,we applied bioinformatic tools to investigate the TERT promoters of Arabidopsis thaliana(L.)Heynh. and other plants,performed the phylogenetic analysis with TERT promoter sequences,and analyzed the TERT gene expression through semi-quantitative RT-PCR.The results showed that the TERT promoter was highly conserved on sequence similarity in the same group of plants.The TERT gene promoter has several regulatory elements related associating to drought,salt and hormone were found.By RT-PCR analysis,the transcription level of At TERT in Arabidopsis Heynh.mutants were always lower than in wild type under normal and stress conditions.In relative to the wild type,the AtTERT mutants became more sensitive under high salt and drought conditions,but no visible difference under abscisic acid(ABA).Taken together,our results veri?ed the prediction of cisacting elements of the AtTERT gene promoter,which might be important in responses to salt and drought stresses.The preliminary result suggestedily proved that the AtTERT promoter regulates the AtTERT gene transcription expression of AtTERT,providing an important reference for the future research on revealing the expression mechanism of this gene.
关 键 词:拟南芥 端粒酶逆转录酶启动子 非生物胁迫 非端粒功能
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