雄蕊特异表达启动子CA55的克隆及其序列分析  

Cloning and Sequence Analysis of Stamen-specific Expression Promoter CA55 from Zea mays L.

作　　者：王纬[1] 黄海泉[1] 张洪波[1] 禹崇惠[1] 黄美娟[1] 

机构地区：[1]西南林业大学园林学院,云南昆明650224

出　　处：《云南农业大学学报（自然科学版）》2013年第3期372-375,共4页Journal of Yunnan Agricultural University:Natural Science

基　　金：云南省自然科学基金项目(2010ZC265);西南林业大学大学生创新性实验计划项目(501601);云南省园林植物与观赏园艺省级重点学科;云南省高校园林植物与观赏园艺重点实验项目

摘　　要：为了调控目的基因仅在雄蕊中表达,本研究采用SDS法提取玉米叶片总DNA,根据雄蕊特异表达启动子的相关序列设计并合成一对引物,通过PCR扩增得到一特异片段,并连接至pMD18-T Vector上进行克隆测序。结果表明该片段全长1 179 bp,与相关序列存在5个碱基的差异,同源性达99.57%,且该片段含有该片段含有2个TATA-box,1个启动序列元件、2个CAAT-box,1个Quantitative-element,8个GTGA-box,1个TACPyAT-box,1个POLLEN1以及3个pollen-box等相关元件,已具备了该启动子所必需的所有调控元件,从而为后期雄蕊特异表达的植物载体构建及遗传转化奠定了基础。In order to regulate the expression of target gene only in the stamen, genomic DNA from Zea mays L. was isolated by using modified SDS method in this study. A pair of specific primers was de signed according to the sequences of stamen specific expression promoters reported in Genbank. A spe cific fragment was amplified by PCR method, ligated into the pMD18T vector and sequenced. The re sults showed that the obtained fragment contained 1 179 bp, that there were five different bases between the sequence tested in this study and the one reported in Genbank, and the homology of their nucleotide sequences reached 99. 57% ; that it contained two TATAbox, one initiator, two CAATbox, one pollen quantitative element, eight GTGAbox, one TACPyATbox, one POLLEN1 and three pollenbox, which possessed all the necessary elements of this kind of promoter. It laid the foundation for the future part of the construction of plant expression vector using this promoter and its genetic transformation.

关 键 词：玉米 雄蕊特异表达启动子 CA55 基因克隆 序列分析 

分 类 号：S603[农业科学—园艺学]