紫草LePS-2基因启动子的克隆及序列分析

Cloning and Sequence Analysis of the Promoter of LePS-2 Gene in Lithospermum erythrorhizon

目的克隆紫草LePS-2基因的启动子,分析该启动子的调控元件,为研究LePS-2基因的表达调控机制奠定基础。方法以紫草愈伤组织基因组DNA为模板,利用Genome walking技术扩增LePS-2基因上游启动子序列,将序列提交至PLACE网站对该启动子进行调控元件预测。结果获得LePS-2基因启动子区域序列1164bp,PLACE软件分析表明,该启动子中存在根特异性元件、暗诱导性元件、激素应答与调节相关元件以及一些转录因子结合位点等。结论启动子元件ATATT和YTCANTYY可能分别赋予LePS-2表达的根特异性和暗诱导性;另外在LePS-2基因启动区域还发现MJ应答元件TGACG,表明MJ有可能参与对LePS-2的表达调控。

OBJECTIVE To clone the promoter of LePS-2 gene of Lithospermum erythrorhizon,and to analyze its mechanism of the expression regulation.METHODS 1164bp of LePS-2 promoter fragment was amplified from Lithospermum callus genomic DNA by using Genome Walking method,which was then submitted to the PLACE website for regulatory element predict.RESULTS 1164bp of LePS-2 promoter fragment was obtained;and several important cis-acting elements,including root organ-specific elements,dark-induced elements,regulatory elements for hormones as well as some transcription factor binding sites were recognized by PLACE program.CONCLUSION Motif-ATATT and motif-YTCANTYY may respectively confer root-specific expression and dark-induced pattern of LePS-2.Furthermore,a MJ response element ＂TGACG＂ was found in the promoter region,which shows that MJ may participate in regulating LePS-2 gene expression.