小麦avenin—like type—B基因启动子的分离及表达载体的构建

根据已知的小麦ALPtype-B基因序列,采用反向PCR技术,从小麦品种鄂恩1号(En1)中克隆得到了ALPtype-B基因的上游启动子序列1664bp。通过PLACE和plantCARE等数据库对所克隆的启动子进行生物信息学分析,发现该启动子除具备启动子的基本元件如TATA—box、CAAT—box等外,还含有胚乳特异性表达的特有元件如Endospermmotif、GCN4-1ikemotif(GLM)、RYmotif和G—box等。用ALPtype—B启动子置换质粒pBll21上的35S启动子,将其与gus基因连接构建植物表达载体。...

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Published in广东农业科学 Vol. 39; no. 4; pp. 1 - 4
Main Author 宋斐 陈泠 孙杨柳 王菲菲 汪越胜 杨广笑 何光源
Format Journal Article
LanguageChinese
Published 湖北省农科院粮食作物研究所,湖北武汉430064 2012
华中科技大学生命科学与技术学院中英HUST-Rres基因工程和基因组学联合实验室/科技部国际科技合作基地(基因工程)/教育部分子生物物理重点实验室,湖北武汉,430074%华中科技大学生命科学与技术学院中英HUST-Rres基因工程和基因组学联合实验室/科技部国际科技合作基地(基因工程)/教育部分子生物物理重点实验室,湖北武汉430074
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ISSN1004-874X
DOI10.3969/j.issn.1004-874X.2012.04.001

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Summary:根据已知的小麦ALPtype-B基因序列,采用反向PCR技术,从小麦品种鄂恩1号(En1)中克隆得到了ALPtype-B基因的上游启动子序列1664bp。通过PLACE和plantCARE等数据库对所克隆的启动子进行生物信息学分析,发现该启动子除具备启动子的基本元件如TATA—box、CAAT—box等外,还含有胚乳特异性表达的特有元件如Endospermmotif、GCN4-1ikemotif(GLM)、RYmotif和G—box等。用ALPtype—B启动子置换质粒pBll21上的35S启动子,将其与gus基因连接构建植物表达载体。
Bibliography:SONG Fei,CHEN Ling,SUN Yang-liu,WANG Fei-fei,WANG Yue-sheng, YANG Guang-xiao,HE Guang-yuan (1.China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory~The Genetic Engineering International Cooperation Base of Chinese Ministry of Science and Technology~Key Laboratory of Molecular Biophysics of Chinese Ministry of Education, College of Life Science and Technology, Huazhong University of Science & Technology (HUST),Wuhan 430074,China; 2.1nstitute of Food Crops,Hubei Academy of A gricultural Sciences, Wuhan 430064,China)
44-1267/S
The study successfully cloned a 1 664 bp upsteam sequence of ALP from wheat Enl using Inverse-PCR (IPCR),based on the sequence of ALP type-B gene.Putative fuctional promoter elements were analyzed by the PLACE datebase and PlantCARE datebase. The results showed that the upsteam sequence contained the basic elements such as CAAT-box and TATA-box. Endosperm motif, GCN4- like motif (GLM),RY motif and G-box were also detected as endosperm-specific elements.Expression vector harbor
ISSN:1004-874X
DOI:10.3969/j.issn.1004-874X.2012.04.001