摘要
为获得在早期胚中特异性表达启动子,本研究通过基因组步移技术对AhDGAT3(GenBank:AY875644.1)的启动子进行克隆。研究结果表明,获得了AhDGAT3起始密码子ATG上游5'侧翼序列2 181bp,应用PLACE和plantCARE在线分析显示,AhDGAT3启动子除了含有核心调控元件TATA-box和CAAT-box之外,还有多个非生物胁迫作用元件,如茉莉酸响应元件、防御和干旱胁迫响应元件TC-rich repeats、光响应元件、干旱诱导的MYB结合位点、光响应MYB结合位点等,以及种子表达相关顺式作用元件及分裂组织表达相关元件。构建重组载体p BI121-PAhDGAT3,转化拟南芥,GUS染色结果显示,该启动子能驱动下游GUS基因,仅在发育早期的心型胚期至鱼雷型胚期较短时间内的胚中特异性表达。
To characterize an early embryo-specific promoter,the putative promoter region of AhDGAT3（ GenBank： AY875644. 1） was isolated by nested PCR-based genomic walking method. Results showed that the 2181 bp 5＇ flanking sequence of AhDGAT3 was isolated. Sequence analysis in PLACE and plant CARE database identified the core regulatory elements such as TATA-box and CAAT-box,the putative cis-acting regulatory elements involved in Me JA-responsiveness,defense stress responsiveness,light-responsiveness,heat stress responsiveness and further the seed or embryo-specific elments. For the functional and characteristic analysis of AhDGAT3 promoter,the recombinant vector p BI121-PAhDGAT3 was designated and transformed into Arabidopsis. Histochemical staining revealed that the activity of p AhDGAT3 was restricted to a short temporal and spatial window from the heart stage to torpedo stage of the embryo.
出处
《中国油料作物学报》
CAS
CSCD
北大核心
2018年第1期25-34,共10页
Chinese Journal of Oil Crop Sciences
基金
国家高技术研究与发展计划(863计划)(2013AA102602-6)
河南省重大科技专项(161100111000)
国家现代农业产业技术体系项目(CARS-13)
河南省现代农业产业技术体系项目(S2012-5)
关键词
花生
DGAT3
早期胚
种子特异性启动子
GUS报告基因
peanut （Arachis hypogaea L. ）
DGAT3
early embryo
seed - specific promoter
GUS reporter gene
