拟南芥菜花药绒毡层启动子的克隆和序列分析  

Isolation and Sequencing Analysis of the Promoter for an Anther specific Gene from Arabidopsis thaliana

作　　者：刘大文[1] 谢友菊[1] 王守才[1] 戴景瑞[1] 

机构地区：[1]中国农业大学,北京100094

出　　处：《作物学报》2000年第4期406-410,共5页Acta Agronomica Sinica

基　　金：国家"九五"攻关项目支持! (96 - 0 0 2 - 0 2 - 0 4)

摘　　要：以拟南芥菜 ( Arabidopsis thaliana)基因组 DNA为模板 ,通过 PCR扩增得到绒毡层特异表达基因 A9的启动子片段 ,克隆到 p UC18载体上。序列分析表明 ,该启动子大小为 360 bp,RNA聚合酶识别序列 TATA box,花药特异表达和增强序列 TGTGG、 TGTGA两个 Motifs皆完整 ,与已报道的序列比较仅有 3个核苷酸发生改变 ,同源性为 99.2 %。该启动子与 GUS基因融合后用基因枪转化玉米花药 ,荧光分析显示花药中有 GUS酶活性存在。The upstream regulatory region of the tapetal specific gene A9 was amplified from Arabidopsis thaliana genome by polymerase chain reaction and cloned into HincⅡ and KpnⅠ site of pUC18. Sequence analysis showed that the cloned fragment contained 364 nucleotides, and shared a sequence homology of 99.2% with the reported A 9 promoter. The putative TATA box was present at position -76 to -69. Two motifs, TGTGG and TGTGA, which were considered to be responsible for the specificity and enhancement of gene expression in the tapetal, were found within -291 to -287 and -243 to -239 region respectively. Gene construct that contained the β glucuronidase ( GUS ) gene under the control of A 9 promoter or CaMV 35 s promoter was introduced into anthers and calli of maize by particle bombardmend. The GUS activity was detected only in anthers by fluorometric assay.

关 键 词：拟南芥菜 启动子 花药特异表达 遗传转化 克隆 

分 类 号：Q943[生物学—植物学]