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拟南芥AtNUDT2启动子的分离及其功能分析

Cloning and Functional Analysis of Promoter AtNUDT2 in Arabidopsis thaliana
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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pNUDT2P-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Pst.DC3000AvrB对该启动子没有诱导作用。 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.
作者 张秀春 吴坤鑫 孙建波 夏亦荠 彭明 李文彬
机构地区 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 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
出处 《热带生物学报》 2012年第2期116-120,共5页 Journal of Tropical Biology
基金 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) 热带生物技术研究所(ITBBZX0811)项目资助
关键词 启动子 AtNUDT2 克隆 GUS染色 promoter AtNUDT2 clone GUS staining
分类号 Q785 [生物学—分子生物学]
  • 相关文献

参考文献17

  • 1MCLENNAN A G. The Nudix hydrolase superfamily[J]. Cell Mol Life Sci, 2006, 63(2) : 123 -143.
  • 2BESSMAN M J, FRICK D N, O' HANDLEY S F. The MutT proteins or "Nudix" hydrolases, a family of versatile, widely dis- tributed, "housecleaning" enzymes [ J ]. J Biol Chem. 1996, 271 (41) : 25059 - 25062.
  • 3MCLENNAN A G, CARTWRIGHT J L, GASMI L. The human NUDT family of nucleotide hydrolases Enzymes of diverse sub- strate specificity[ J]. Adv Exp Med Biol,2000,486:115 -118.
  • 4OGAWA T, UEDA Y, YOSHIMURA K, et al. Comprehensive analysis of cytosolic Nudix hydrolases in Arabidopsis thaliana [J]. Biol Chem,2005 ,280(26) : 25277 -25283.
  • 5KLAUS S M, WEGKAMP A, SYBESMA W, et al. A Nudix enzyme removes pyrophosphate from dihydroneopterin triphos- phate in the folate synthesis pathway of bacteria and plants[J]. J Biol Chem , 2005 ,280(7) : 5274 -5280.
  • 6YOSHIMURA K, OGAWA T, UEDA Y, et al. At-NUDX1, an 8-oxo-7,8-dihydro-2' -deoxyguanosine 5' -triphosphate pyro- phosphohydrolase, is responsible foreliminating oxidized nucleotides in Arabidopsi- [ J ]. Plant Cell Physiol, 2007,48 : 1438 - 1449.
  • 7OLEJNIK K, KRASZEWSKA E. Cloning and characterization of an Arabido_psis thaliana Nudix hydrolase homologous to the mammalian GFG protein [ J ]. Bioehim Biophys Aeta. 2005,1752 ( 2 ) : 133 - 141.
  • 8JAMBUNATHAN N, MAHALINGAM R. Analysis of Arabidopsis Growth Factor Gene 1 ( GFG1 ) encoding a Nudix hydrolase during oxidative signaling [ J ]. Planta, 2005 ( 3 ) : 1 - 11.
  • 9BARTSCH M, GOBBATO E, BEDNAREK P, et al. Salicylic acid-independent ENHANCED DISEASE SUSCEPTIBILITY1 signaling in Arabidopsis immunity and cell death is regulated by the monooxygenase FMO1 and the Nudix hydrolase NUDT'/ [J]. Plant Cell. 2006, 18:1038-1051.
  • 10OGAWA T, ISHIKAWA K, HATADA K, et al. Overexpression of an ADP-ribose pyrophosphatase, AtNUDX2, confers en- hanced tolerance to oxidative stress in Arabidopsis plants[J]. The Plant Journal, 2009,57:289 -301.

二级参考文献12

  • 1MCLENNAN A G. The Nudix hydrolase superfamily[ J]. Cell Mol Life Sci. 2006,63 (2) :123 -143.
  • 2BESSMAN M J, Frick D N, O' HANDLEY S F. The MutT proteins or "Nudix" hydrolases, a family of versatile, widely dis- tributed, "housecleaning" enzymes [ J ]. J Biol Chem. 1996,271 (41 ) :25059 - 25062.
  • 3MCLENNAN A G, CARTWRIGHT J L, GASMI L. The human NUDT family of nucleotide hydrolases. Enzymes of diverse substrate specificity[ J]. Adv Exp Med Biol. 2000,486 : 115 - 118.
  • 4OGAWA T, UEDA Y, YOSHIMURA K, et al. Comprehensive analysis of eytosolic Nudix hydrolases in Arabidopsis thaliana [ J ]. J Biol Chem. 2005,280 (26) :25277 - 25283.
  • 5KLAUS S M, WEGKAMP A, SYBESMA W, et al. A nudix enzyme removes pyrophosphate from dihydroneopterin triphosphate in the folate synthesis pathway of bacteria and plants[J].J Biol Chem. 2005,280(7) :5274 -5280.
  • 6YOSHIMURA K, OGAWA T, UEDA Y, et al. At-NUDX1, an 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate pyro- phosphohydrolase, is responsible foreliminating oxidized nucleotides in Arabidopsis[ J]. Plant Cell Physiol. 2007,48:1438 - 1449.
  • 7OLEJNIK K, KRASZEWSKA E. Cloning and charactarization of an Arabidopsis thaliana Nudix hydrolase homologous to the mammalian GFG protein [ J ]. Biochim Biophys Acta. 2005,1752 (2) :133 -141.
  • 8JAMBUNATHAN N, MAHALINGAM R. Analysis of Arabidopsis Growth Factor Gene 1 ( GFG1 ) encoding a nudix hydrolase during oxidative signaling[ J]. Planta. 2005 ( 3 ) : 1 - 11.
  • 9BARTSCH M, GOBBATO E, BEDNAREK P, et al. Salicylic Acid-Independent ENHANCED DISEASE SUSCEPTIBILITY1 Signaling in Arabidopsis Immunity and Cell Death Is Regulated by the Monooxygenase FMO1 and the Nudix Hydrolase NUDT7 [ J]. Plant Cell. 2006,18 : 1038 - 1051.
  • 10OGAWA T, ISHIKAWA K, HARADA K, et al. Overexpression of an ADP-ribose pyrophosphatase, AtNUDX2, confers en- hanced tolerance to oxidative stress in Arabidopsis plants[J]. The Plant Journal. 2009,57:289- 301.

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热带生物学报
2012年 第2期

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