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Volume 3 Issue 2
Jun.  2012
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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. doi: 10.15886/j.cnki.rdswxb.2012.02.001
Citation: 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. doi: 10.15886/j.cnki.rdswxb.2012.02.001

Cloning and Functional Analysis of Promoter AtNUDT2 in Arabidopsis thaliana

doi: 10.15886/j.cnki.rdswxb.2012.02.001
  • Received Date: 2010-12-29
  • 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.thaliana through Agrobacterium tumefaciens by mediating.A batch of transgenic A.thaliana 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.
  • [1] MCLENNAN A G.The Nudix hydrolase superfamily[J].Cell Mol Life Sci,2006,63(2):123-143.
    [2] BESSMAN 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.
    [3] MCLENNAN 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.
    [4] OGAWA T,UEDA Y,YOSHIMURA K,et al.Comprehensive analysis of cytosolic Nudix hydrolases in Arabidopsis thaliana[J].Biol Chem,2005,280(26):25277-25283.
    [5] KLAUS 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.
    [6] YOSHIMURA K,OGAWA T,UEDA Y,et al.At-NUDX1,an8-oxo-7,8-dihydro-2’-deoxyguanosine5’-triphosphate pyro-phosphohydrolase,is responsible foreliminating oxidized nucleotides in Arabidopsis[J].Plant Cell Physiol,2007,48:1438-1449.
    [7] OLEJNIK K,KRASZEWSKA E.Cloning and characterization of an Arabidopsis thaliana Nudix hydrolase homologous to the mammalian GFG protein[J].Biochim Biophys Acta.2005,1752(2):133-141.
    [8] JAMBUNATHAN N,MAHALINGAM R.Analysis of Arabidopsis Growth Factor Gene1(GFG1)encoding a Nudix hydrolase during oxidative signaling[J].Planta,2005(3):1-11.
    [9] BARTSCH M,GOBBATO E,BEDNAREK P,et al.Salicylic acid-independent ENHANCED DISEASE SUSCEPTIBILITY1signaling in Arabidopsis immunity and cell death is regulated by the monooxygenase FMO1and the Nudix hydrolase NUDT7[J].Plant Cell.2006,18:1038-1051.
    [10] OGAWA 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.
    [11] DE BLOCK M,VERDUYN C,DE BROUWER,et al.Poly(ADP-ribose)polymerase in plants affects energy homeostasis,cell death and stress tolerance[J].Plant J,2005,41:95-106.
    [12] 张秀春,李文彬,夏亦荠,等.AtNUDT8过量表达的拟南芥转基因植株[J].热带生物学报,2010,1(1):8-11.
    [13] WHALEN M C,INNES R W,BENT A F,et al.Identification of pseudomonas syringae pathogens of Arabidopsis and a bac-terial locus determining avirulence on both Arabidopsis and soybean[J].Plant Cell,1991,3(1):49-59.
    [14] JEFFERSON R A,KAVANAGH T A,BEVAN M W.GUS fusions:β-glucuronidase as a sensitive and versatile gene fusion marker in higherplants[J].EMBO J,1987(6):3901-3907.
    [15] JAMBUNATHAN N,PENAGANTI A,TANG Y,et al.Modulation of redox homeostasis under suboptimal conditions by Ara-bidopsis Nudix hydrolase7[J].BMC Plant Biol,2010,10:173.
    [16] ADAMS-PHILLIPS L,WAN J,TAN X,et al.Discovery of ADP-ribosylation and other plant defense pathway elements through expression profiling of four different Arabidopsis-Pseudomonas R-avr interactions[J].Mol Plant Microbe Interact,2008,21(5):646-57.
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Cloning and Functional Analysis of Promoter AtNUDT2 in Arabidopsis thaliana

doi: 10.15886/j.cnki.rdswxb.2012.02.001

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.thaliana through Agrobacterium tumefaciens by mediating.A batch of transgenic A.thaliana 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.

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. doi: 10.15886/j.cnki.rdswxb.2012.02.001
Citation: 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. doi: 10.15886/j.cnki.rdswxb.2012.02.001
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