摘要
为了解文心兰生物钟基因OnELF3的转录调控,本研究采用TAIL-PCR技术从文心兰基因组中克隆到OnELF3基因起始密码子上游2 204 bp的启动子序列。使用BDGP、PlantCARE和PLACE在线软件对OnELF3基因启动子的转录起始位点与顺式作用元件进行预测。结果表明启动子序列除包含TATA-box和CAAT-box等启动子基本元件外,还包含组织特异性元件、光调控元件、植物激素响应元件、胁迫反应响应元件和昼夜节律调控元件等。为探究OnELF3启动子的表达活性,构建pCAMBIA1301-p OnELF3p:GUS载体,利用农杆菌介导法,转化烟草与拟南芥。烟草叶片瞬时转化表明克隆的OnELF3启动子序列具有启动子活性。转化拟南芥结果表明,OnELF3启动子能够驱动下游的GUS基因在T2代拟南芥中稳定表达,GUS组织染色显示该启动子呈现发育与组织特异性表达。这些结果为进一步研究文心兰OnELF3基因的转录表达调控与相关功能分析提供基础。
In order to understand the transcriptional regulation of Oncidium circadian-clock gene OnELF3,the2 204 bp promoter sequence of the OnELF3 gene was cloned from Oncidium genomic DNA by the TAIL-PCR technique in this research.Prediction of the transcription initiation sites(TSSs) and the cis-acting elements of the OnELF3 gene promoter were conducted with the BDGP,PlantCARE and PLACE online software.The results showed that the promoter sequence contains the basic elements of TATA-box and CAAT-box,as well as other promoter elements such as tissue-specific elements,light regulation response elements,hormone response responsive elements,stressresponse elements and circadian rhythm regulatory elements.In order to explore the expression activity of the promoter,the pCAMBIA1301-p OnELF3 p:GUS vector was constructed and transformed into tobacco and Arabidopsis thaliana with Agrobacterium-mediated method.The transient transformation of tobacco leaves indicated that the cloned OnELF3 promoter sequence has promoter activity.The transformation of Arabidopsis showed that the OnELF3 promoter could drive the downstream GUS gene to stably express in T2 Arabidopsis plants.GUS histoch emical staining demonstrated a developmental and tissue-specific expression pattern of the OnELF3 promoter.These results lay a foundation for further study on the transcriptional regulation and related functional analysis provides the basis of Oncidium OnELF3 gene.
作者
谭玉荣
高璇
王丹
张恒
李双江
王鹏
李晨
徐丹
李梦桃
潘英文
刘进平
Tan Yurong;Gao Xuan;Wang Dan;Zhang Heng;Li Shuangiang;Wang Peng;Li Chen;Xu Dan;Li Mengtao;Pan Yingwen;Liu Jinping(Hainan Key Laboratory for Sustainable Uilzaion of Tropical Bioresources,Cllge of Tropical Crops,Hainan Univrsity,Haikou,570228;Post-Em-try Quarantine Statio for Tropical Plant,Hainan Entry-Exit Inspection and Quarantine Bureau,Haikou,570311)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第15期4898-4907,共10页
Molecular Plant Breeding
基金
海南省基础与应用基础研究计划(自然科学领域)高层次人才项目(2019RC147)
国家自然科学基金项目(31560573)
海南省研究生创新科研课题(Hys2019140
Hyb2019-14)
海南大学作物学科研究生科研创新课题(ZWCX2018030)共同资助。
