麻疯树异戊烯基焦磷酸异构酶基因(IPI)启动子的克隆及瞬时表达分析  

Cloning and transient expression analysis of isopentenyl diphosphate isomerase gene (IPI) promoter from Jatropha curcas

作　　者：胡晓乐[1] 张敏[1] 田银帅[1] 徐莺[1] 陈放[1] HU Xiao-Le ZHANG Min TIAN gin-Shuai XU Ying CHEN Fang(College of Life Sciences, Sichuan University, Chengdu 610064, China)

机构地区：[1]四川大学生命科学学院,成都610064

出　　处：《四川大学学报（自然科学版）》2016年第6期1409-1414,共6页Journal of Sichuan University(Natural Science Edition)

基　　金：国家"十二五"重大科技支撑计划课题(2011BAD22B08)

摘　　要：本文以麻疯树基因组DNA为模板,通过PCR扩增得到麻疯树异戊烯基焦磷酸异构酶(IPI)基因(JcIPI)起始密码子上游1 536bp的启动子序列.利用在线软件PLACE和PlantCARE分析表明该序列除具备TATA-Box、CAAT-Box等启动子基本元件外,还含有AuxRR-core、TATC-box、MBS、HSE等特异性元件.为确定启动子核心启动区域,构建JcIPI启动子283、550、970、1 276和1 536bp的5′端缺失片段,并将其分别驱动pBI121载体的葡萄糖苷酸酶(GUS)基因,构建植物表达载体.采用农杆菌介导法转化烟草,在烟草叶片中进行瞬时表达分析.GUS酶活测定结果显示,五个启动子缺失片段都具有启动子活性,并随长度增加而增强.In order to study the expression and regulation of isopentenyl diphosphate isomerase （IPI） gene in the terpenoid biosynthesis pathway of Jatropha curcas, a 1536bp promoter fragment of IPI gene was cloned by PCR method. The bioinformatics analysis showed that the fragment contained the con- served promoter sequence, such as TATA-box, CAAT-box. Furthermore, it contained several elements related to auxin response, gibberellin response, disease-related and high temperature. To determine the optimal promoter sequence for gene expression, IPI gene promoter was deleted from its 5＇ end to form promoter fragments with 283, 550, 970, 1276 and 1536bp. Such fragments were fused to a 13-glucuroni- dase （GUS） gene. The fused genes were transformed into Nicotiana benthamiana using Agrobacterium tumefaeiens-mediated method for transient expression. GUS quantitative fluorometric assays demonstra- ted that the five different length fragments of the promoter had promoter activities and the activities were increased with the deleted length of promoters.

关 键 词：麻疯树 异戊烯基焦磷酸异构酶 启动子 瞬时表达 

分 类 号：Q78[生物学—分子生物学]