拟南芥AtNUDT10启动子的分离及其功能分析  

Cloning and Function Analysis of Promoter of AtNUDT10 in Arabidopsis thaliana

作　　者：张秀春[1,2] 李若霖[1,3] 唐文[1,3] 夏亦荠[1] 彭明[1,2] 吴坤鑫[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所/农业部热带作物生物学与遗传资源利用重点实验室,海口571101 [2]农业部转基因植物及植物用微生物环境安全监督检验测试中心,海口571101 [3]海南大学农学院,海口570228

出　　处：《中国农学通报》2012年第21期164-168,共5页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金(30760196);中央级公益性科研院所基本科研业务费专项资金(中国热带农业科学院院本级)资助项目(1630052012007)

摘　　要：根据已发表的拟南芥基因组序列设计合成一对引物,以拟南芥基因组DNA为模板克隆AtNUDT10上游非编码区,并与pBAR-GUS3相连构建了植物表达载体pNUDT10P-GUS,采用农杆菌GV3101介导的渗透法转化野生型拟南芥,通过除草剂筛选获得了一批抗性纯合子转基因植株。然后对转基因植株2周幼苗进行GUS活性的组织化学染色分析,并对3.5～4周的转基因植株叶片进行病原菌诱导表达分析。结果表明,已获得AtNUDT10启动子,该启动子为组成型表达的启动子,并且病原菌Pst.DC3000及Pst.DC3000AvrB对该启动子没有诱导作用。5＇-UTR of AtNUDTIO was cloned by PCR from Arabidopsis thaliana and was inserted into pBAR-GUS to form a recombined construct pNUDTIOP-GUS. The construct was then used to transform wide-type Arabidopsis thaliana by Agrobacterium tumefaciens mediating. A batch of transgenic Arabidopsis thaliana with 5＇-UTR of AtNUDT10 was obtained through glufosinate screening. The results showed that, we had got the promoter of AtNUDT2 that was constitutive promoter. Furthermore, the expression of the promoter could not be induced by the Pst.DC3000 and Pst.DC3000 AvrB.

关 键 词：启动子 AtNUDT10 克隆 GUS染色 

分 类 号：S685.12[农业科学—观赏园艺]