| 拟南芥AtNUDT2启动子的分离及其功能分析 |
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| 引用本文： | 张秀春,吴坤鑫,孙建波,夏亦荠,彭明,李文彬.拟南芥AtNUDT2启动子的分离及其功能分析[J].热带生物学报,2012,3(2):116-120. |
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| 作者姓名： | 张秀春 吴坤鑫 孙建波 夏亦荠 彭明 李文彬 |
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| 作者单位： | [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,海南海口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,海南海口571101 |
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| 基金项目： | 国家自然科学基金（30760196,31070608）;中央级公益性科研院所基本科研业务专项资金,中国热带农业科学院院本级（1630052012007）及热带生物技术研究所（ITBBZX0811）项目资助 |
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| 摘 要： | 根据已发表的拟南芥基因组序列设计合成1对引物，以拟南芥基因组DNA为模板克隆AtNUDT2上游非编码区，并将其与pBAR－GUS3相连，构建了植物表达载体pNUD12P－GUS，采用以农杆菌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AtNUDT2启动子，且该启动子为组成型表达启动子；病原菌Pst．DC3000及风t．DC3000 AvrB对该启动子没有诱导作用。
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| 关 键 词： | 启动子 AtNUDT2 克隆 GUS染色 |
Cloning and Functional Analysis of Promoter AtNUDT2
in Arabidopsis thaliana |
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| ZHANG Xiu-chun,WU Kun-xin,SUN Jian-bo,XIA Yi-ji,PENG Ming,LI Wen-bin.Cloning and Functional Analysis of Promoter AtNUDT2
in Arabidopsis thaliana[J].Journal of Tropical Biology,2012,3(2):116-120. |
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| Authors: | ZHANG Xiu-chun WU Kun-xin SUN Jian-bo XIA Yi-ji PENG Ming LI Wen-bin |
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| Institution: | 1. Institute of Tropical Bioscienee and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, and Key laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, I-Iaikou 571101 ,China; 2. Environmental Safety Supervision and Inspection Centre for Genetically Modified Plants and Microorganisms used in Plants, Ministry of Agriculture, Haikou 571101, China) |
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| Abstract: | 5＇ -UTR of AtNUDT2 was cloned by PCR from Arabidopsis thaliana and inserted into pBAR-GUS to form a recombined construct pNUDT2P-GUS. The construct was then used to transform wide-type A. t~ through Agrobacterium tumefaciens by mediating. A batch of transgenic A. thal/ana with 5 ＇ -UTR of AtNUDT2 was obtained through glufosinate screening. The transgenic plants （2 weeks old） were then histochemically stained to assess their GUS activity, and the leaves of the 3.5 - 4 weeks old plants were inoculated with pathogens to observe their pathogenic expression. The results showed the promoter of AtNUDT2 produced is constitutive. Furthermore, the expression of the promoter were not induced by the Pst. DC3000 and Pst. DC3000 AvrB. |
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| Keywords: | promoter AtNUDT2 clone GUS staining |
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