甘薯块根储藏蛋白(Sporamin)启动子克隆及功能验证  

Cloning and Sequence Analysis of the Promoter of Sporamin Gene from Sweetpotato(Ipomoea batatas)

作　　者：张聪[1] 郑雪莲[3] 蒲志刚[1] 张婷[1] 吴洁[1] 谭文芳[2] 王大一[2] 阎文昭[1] 

机构地区：[1]四川省农业科学院生物技术核技术研究所,成都610066 [2]四川省农业科学院作物研究所,成都610066 [3]电子科技大学生命科学学院,成都610054

出　　处：《分子植物育种》2012年第6期707-713,共7页Molecular Plant Breeding

基　　金：国家现代农业甘薯产业技术体系建设专项资金(CARS-11-B-04);四川省农作物育种攻关项目甘薯育种子课题(2011-2015);四川省基因工程育种项目(2011-2015);国家自然科学基金项目(31000735)共同资助

摘　　要：提取甘薯"川薯34"总DNA,设计引物,通过高保真PCR扩增获得了长为1144bp的甘薯储藏蛋白(Sporamin)基因启动子SpoA-p。PLACE在线分析表明:Sporamin启动子具有基本的启动子元件CAAT-box,并包含大量与储藏蛋白表达相关、抗逆相关及蔗糖表达相关的功能组件。构建植物表达载体pBI-SpoA-p::GUS、农杆菌工程菌株EHA105:pBI-SpoA-p::GUS后导入烟草及甘薯,分别得到15株烟草和10株甘薯的阳性转基因植株,GUS染色分析表明烟草各部位中只在根中有基因表达,甘薯根、茎和叶各部位未检测到活性。5%蔗糖诱导处理24h后,在烟草根、茎及叶均观察到GUS活性;在甘薯中只有根有GUS活性。因此,可以看出此启动子为甘薯根部特异的表达启动子,并受蔗糖诱导而表达。A 1 100 bp DNA fragment named SpoA-pro was amplified from a sweet potato cultivar（Chuanshu34）,by PCR with high-fidelity DNA polymerase KOD-plus.PLACE online analysis revealed that the promoter of Sporamin contains basic cis-elements of promoter such as CAAT-box,and some functional domains associated with the expression of storage protein,stress response and sucrose inducible in the sequence.Furthermore,the plant expression vector for this promoter was constructed,and then transformed in tobacco and sweet potato mediated by Agrobacterium.15 tobacco transgenic plants and 18 sweet potato transgenic plants were obtained and identified by PCR analysis,GUS staining assay showed blue color only happens in roots of transgenic tobacco,whereas the blue color appears in roots,stems and leaves of transgenic sweet potato.While transgenic the plants were treated with 5% sucrose,strong GUS staining were detected in the roots,leaves and stems of transgenic tobacco butonly in roots of transgenic sweet potato.Therefore,we thought that the SpoA-pro smight be a promoter that should have the characteristics of root specific and sucrose-inducible expression.

关 键 词：甘薯 储藏蛋白基因 启动子 植物表达载体 蔗糖诱导 根部特异的表达 

分 类 号：S531[农业科学—作物学]