• Title/Summary/Keyword: Promoter activation

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Molecular Cloning and Functional Analysis of Rice (Oryza sativa L.) OsNDR1 on Defense Signaling Pathway

  • Lee, Joo-Hee;Kim, Sun-Hyung;Jung, Young-Ho;Kim, Jung-A;Lee, Mi-Ok;Choi, Pil-Gyu;Choi, Woo-Bong;Kim, Kyung-Nam;Jwa, Nam-Soo
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.149-157
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    • 2005
  • A novel rice (Oryza sativa L.) gene, homologous to Arabidopsis pathogenesis-related NDR1 gene, was cloned from cDNA library prepared from 30 min Magnaporthe grisea -treated rice seedling leaves, and named as OsNDR1. OsNDR1 encoded a 220-aminoacid polypeptide and was highly similar to the Arabidopsis AtNDR1 protein. OsNDR1 is a plasma membrane (PM)-localized protein, and presumes through sequence analysis and protein localization experiment. Overexpression of OsNDR1 promotes the expression of PBZ1 that is essential for the activation of defense/stressrelated gene. The OsNDR1 promoter did not respond significantly to treatments with either SA, PBZ, or ETP. Exogenously applied BTH induces the same set of SAR genes as biological induction, providing further evidence for BTH as a signal. Presumably, BTH is bound by a receptor and the binding triggers a signal transduction cascade that has an ultimate effect on transcription factors that regulate SAR gene expression. Thus OsNDR1 may act as a transducer of pathogen signals and/or interact with the pathogen and is indeed another important step in clarifying the component participating in the defense response pathways in rice.

Isolation and Characterization of Pathogen-Inducible Putative Zinc Finger DNA Binding Protein from Hot Pepper Capsicum annuum L.

  • Oh, Sang-Keun;Park, Jeong-Mee;Jung, Young-Hee;Lee, Sanghyeob;Kim, Soo-Yong;Eunsook Chung;Yi, So-Young;Kim, Young-Cheol;Seung, Eun-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.79.2-80
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    • 2003
  • To better understand plant defense responses against pathogen attack, we identified the transcription factor-encoding genes in the hot pepper Capsicum annuum that show altered expression patterns during the hypersensitive response raised by challenge with bacterial pathogens. One of these genes, Ca1244, was characterized further. This gene encodes a plant-specific Type IIIA - zinc finger protein that contains two Cys$_2$His$_2$zinc fingers. Ca1244 expression is rapidly and specifically induced when pepper plants are challenged with bacterial pathogens to which they are resistant. In contrast, challenge with a pathogen to which the plants are susceptible only generates weak Ca1244 expression. Ca1244 expression is also strongly induced in pepper leaves by the exogenous application of ethephon, an ethylene releasing compound. Whereas, salicylic acid and methyl jasmonate had moderate effects. Pepper protoplasts expressing a Ca1244-smGFP fusion protein showed Ca1244 localizes in the nucleus. Transgenic tobacco plants overexpressing Ca1244 driven by the CaMV 355 promoter show increased resistance to challenge with a tobacco-specific bacterial pathogen. These plants also showed constitutive upregulation of the expression of multiple defense-related genes. These observations provide the first evidence that an Type IIIA - zinc finger protein, Ca1244, plays a crucial role in the activation of the pathogen defense response in plants.

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Cloning And Characterization of Pathogen-Inducible EREBP-Like Transcription Factor(CaNR19) From Hot Pepper (Capsicum annuum L.)

  • Yi, So-Young;Kim, Jee-Hyub;Yu, Seung-Hun;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.77.2-78
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    • 2003
  • An EREBP/AP2-type transcription factor (CaPFl) was isolated by DDRT-PCR following inoculation of soybean pustule pathogen Xanthomonas axonopodis pv. glycines Bra which induces HR on pepper leaves. Genomic Southern blot analysis revealed that the CaPFl gene is present as a single copy within the hot pepper genome. The deduced amino acid sequence of CaPFl has two potential nuclear localization signals, a possible acidic activation domain, and an EREBP/AP2 motif that could bind to a conserved cis- element present in promoter region of many stress-induced genes. The mRNA level of CaPFl was induced by both biotic and abiotic stresses. We observed higher-level transcripts in resistance-induced pepper tissues than diseased tissues. Expression of CaPFl is also induced upon various abiotic stresses including ethephon, MeJA, cold stress, drought stress and salt stress treatments. To study the role of CPFI in plant, transgenic Arabidopsis and tobacco plants which express higher level of pepper CaPFl were generated. Global gene expression analysis of transgenic Arabidopsis by cDNA microarray indicated that expression of CaPFl in transgenic plants affect the expression of quite a few GCC box and DRE/CRT box-containing genes. Furthermore, the transgenic Arabidopsis and tobacco plant, expressing CaPFl showed tolerance against freezing temperature and enhanced resistance to Pseudomonas syrnigae pv. tabaci. Taken together, these results indicated that CaPFl is a novel EREBP/AP2 transcription factor in hot pepper plant and it may has a significant role(s) in regulation of biotic and abiotic stresses in plant.

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Enhancement of immunomodulatory activity by liposome-encapsulated natural phosphodiester bond CpG-DNA in a human B cell line

  • Kim, Dong-Bum;Rhee, Jae-Won;Kwon, Sang-Hoon;Kim, Young-Eun;Choi, Soo-Young;Park, Jin-Seu;Lee, Young-Hee;Kwon, Hyung-Joo
    • BMB Reports
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    • v.43 no.4
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    • pp.250-256
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    • 2010
  • Natural phosphodiester bond CpG-DNA that contains immunomodulatory CpG motifs (PO-DNA) upregulates the expression of proinflammatory cytokines and induces an Ag-driven Th1 response in a CG sequence-dependent manner in mice. In humans, only phosphorothioate backbone-modified CpG-DNA (PS-DNA) and not PO-DNA has immunomodulatory activity. In this study, we found that liposome-encapsulated PO-DNA upregulated the expression of human $\beta$-defensin-2 (hBD-2) and major histocompatibility class II molecules (HLA-DRA) in a CG sequence-dependent and liposome- dependent manner in human B cells. Of the three different liposomes, DOTAP has the unique ability to enhance the immunomodulatory activity of PO-DNA. In contrast, HLA-DRA and hBD-2 promoter activation can be induced by liposome-encapsulated PS-DNA in a CG sequence-independent manner, depending on the CpG-DNA species. Our observations demonstrate that, when encapsulated with a proper liposome in the immune system, natural PO-DNA has the potential to be a useful therapy for the regulation of the innate immune response.

The Effect of Sn on Dehydrocyclization of n-Heptane over Pt-Sn/γ-Al2O3 Catalyst (Pt-Sn/γ-Al2O3 촉매상에서 n-Heptane의 탈수소고리화 반응에서 조촉매 주석의 영향)

  • Song, Myeong-Sug;Kim, Moon-Chan;Kim, Kyung-Lim
    • Applied Chemistry for Engineering
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    • v.2 no.4
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    • pp.411-420
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    • 1991
  • The dehydrocyclization of n-heptane was studied over $Pt-Sn/{\gamma}-Al/_2O_3$ catalysts with varying Sn content in a fixed bed continuous flow reactor. The range of experimental conditions was at the temperature between 450 and $550^{\circ}C$, the pressure $20{\times}10^5-50{\times}10^5Pa$, the contact time 0.09 and 0.27 hr and the $H_2/H.C$. mole ratio 10. The conversion and selectivity of dehydrocyclization increased with increasing temperature, but decreased with increasing pressure. When we use Sn as a promoter, the selectivity of dehydrocyclization changesa a little, but the conversion was increased and the selectivity of isomerization increased a lot. The activation energy of dehydrocyclization of n-heptane was 34.5 kcal/mol over 0.6 wt % Pt-0.6 wt % $Sn/{\gamma}-Al_2O_3$.

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Root Colonization and ISR-mediated Anthracnose Disease Control in Cucumber by Strain Enterobacter asburiae B1

  • Bharathkumar, S.;Park, Jin-Woo;Han, Ji-Hee;Park, Kyung-Seok
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.333-343
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    • 2009
  • Here, we show that an endophytic bacterial strain, Enterobacter asburiae B1 exhibits the ability to elicit ISR in cucumber, tobacco and Arabidopsis thaliana. This indicates that strain B1 has a widespread ability to elicit ISR on various host plants. In this study, E. asburiae strain B1 did not show antifungal activity against tested major fungal pathogens, Colletotrichum orbiculare, Botrytis cinerea, Phytophthora capsici, Rhizoctonia solani, and Fusarium oxysporum. Moreover, the siderophore production by E. asburiae strain B1 was observed under in vitro condition. In greenhouse experiments, the root treatment of strain B1 significantly reduced disease severity of cucumber anthracnose caused by fungal pathogen C. orbiculare compared to nontreated control plants. By root treatment of strain B1 more than 50% disease control against anthracnose on cucumber was observed in all greenhouse experiments. Simultaneously, under the greenhouse condition, the soil drench of strain B1 and a chemical inducer benzothiadiazole (BTH) to tobacco plants induced GUS activity which is linked with activation of PR promoter gene. Furthermore, in Arabidopsis thaliana plants the soil drench of strain B1 induced the defense gene expression of PR1 and PDF1.2 related to salicylic acid and jasmonic acid/ethylene signaling pathways, respectively. In this study, for the main focus on root colonization by strain B1 associated with defense responses, bacterial cells of strain B1 was tagged with the gfp gene encoding the green fluorescent protein in order to determine the colonization pattern of strain B1 in cucumber. The gfp-tagged B1 cells were found on root surface and internal colonization in root, stem, and leaf. In addition to this, the scanning electron microscopy observation showed that E. asburiae strain B1 was able to colonized cucumber root surface.

Disease-resistant Transgenic Arabidopsis Carrying the expI Gene from Pectobacterium carotovorum subsp. carotovorum SL940

  • Lee, Joo-Hee;Hong, Ja-Bin;Hong, Sang-Bin;Choi, Min-Seon;Jeong, Ki-Yong;Park, Hyoung-Joon;Hwang, Duk-Ju;Lee, Seung-Don;Ra, Dong-Soo;Heu, Sung-Gi
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.183-190
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    • 2008
  • Plant-cell-wall-degrading enzymes (PCWDEs) of Pectobacterium carotovorum subsp. carotovorum are the key virulence factor in pathogenesis of soft rot disease of vegetables. The production of PCWDEs is controlled in a cell density dependent manner to avoid the premature production of PCWDEs and subsequent activation of plant defense. N-oxoacyl-homoserine lactone (OHL) is essential for quorum sensing in the soft rot pathogen and the expI gene is responsible for OHL production. The ExpI homolog isolated from P. carotovorum subsp. carotovorum SL940 had 94% identity with ExpI of E. carotovora subsp. carotovora scc3193 and 74% identity with Carl of E. carotovora subsp. atroseptica. The transgenic plants that express exp I uner the control of CaMV35S promoter were able to produce diffusible OHL. Transgenic plants producing OHL were very resistant to the infection of P. carotovorum subsp. carotovorum. Since the PR1 gene was strongly induced and NPR1 and NPR4 were induced weakly in transgenic plants compared to the wild type, salicylic acid-dependent pathways is likely involved in the resistance to the soft rot pathogen P. carotovorum subsp. carotovorum in ExpI transgenic plants.

In Vitro Transcription Analyses of Autographa californica Nuclear Polyhedrosis Virus Genes

  • Huh, Nam-Eung
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.183-190
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    • 1994
  • Cell-free extracts prepared from cultured insect cells, Spodoptera. frugiperda, were analyzed for activation of early gene transcription of an insect baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV). The template DNA used for in vitro transcription assays contained promoter sites for the baculovirus genes that have been classified as immediate early ($\alpha$) or early genes. These genes are located in the HindIII-K/Q region of the AcNPV genome. Nuclei isolated from the AcNPV-infected Spodoptera frugiperda cells were also used for in vitro transcription analysis by RNase-mapping the labeled RNA synthesized from in vitro run-on reaction in the isolated nuclei. The genes studied by this technique were p26 and pl0 genes which were classified as delayed early and late gene, respectively. We found that transcription of the genes from the HindIII-K region was accurately initiated and unique in the whole cell extract obtained from uninfected cells, although abundance of the in vitro transcripts was reverse to that of in vivo RNA. With isolated nuclei transcription of the p26 gene was inhibited by $\alpha$-amanitin suggesting that the p26 gene was transcribed by host RNA polymerase II. However, transcription of the pl0 gene in isolated nuclei was not inhibited by $\alpha$-amanitin, but rather stimulated by the inhibitor. We also found that the synthesis of $\alpha$-amanitin-resistant RNA polymerase was begun before 6 hr p.i., the time point at which the onset of viral DNA replication as well as the appearance of a-amanitin-resistant viral transcripts were detected. These studies give us strong evidence to support the previous data that early genes of AcNPV were transcribed by host RNA polymerease III, while transcription of late genes was mediated at least by a novel $\alpha$-amanitin-resistant RNA polymerase.

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Catalytic Oxidation of Methane Using the Manganese Catalysts (망간촉매를 이용한 메탄의 산화반응)

  • Jang, Hyun-Tae;Cha, Wang-Seong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.1
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    • pp.537-544
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    • 2011
  • This work was conducted to investigate the oxidation characteristics of methane having the highest ignition temperature among the other hydrocarbon gases using transition metal catalysts. The catalyst used for methane oxidation was manganese oxide having a various oxidation number, such as MnO, $MnO_2$, $Mn_2O_3$, $Mn_3O_4$, $Mn_4O_5$. The manganese oxide(MnxOy) catalyst is impregnated on $TiO_2$, $Al_2O_3$ for methane oxidation. To enhanced both of activity and life time of catalysts, Ni and Co was used as a promoter. In this study, various co-catalysts were synthesized by using excess wet impregnation method. The effect of reaction temperature and space velocity was measured to calculate the activity of catalysts such as, activation energy of $T_{50}$, and $T_{90}$. The life time of bi-metallic manganese mixture, such as Mn-Co and Mn-Ni catalysts, were increased more 10 % than manganese oxide catalyst, but activity of those was decreased slightly.

T7 RNA Polymerase Is Expressed in Plants in a Nicked but Active Form (T7 RNA polymerase 유전자의 담배식물에서의 발현)

  • Caviedes, Miguel A.;Thornburg, Robert W.;Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.40 no.4
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    • pp.271-276
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    • 1997
  • We have prepared several chimeric constructs containing the bacteriophage T7 RNA polymerase gene under control of the wound-inducible potato proteinase inhibitor II (pin2) promoter and have transformed Nicotiana tabacum plants with these constructs. Southern blot analyses indicate that either one or two copies of the gene constructs are present in the transgenic plants. Northern blot analyses indicate that mRNA encoding T7 RNA polymerase is expressed in a wound-inducible manner. We purified T7 RNA polymerase and prepared antiserum. This antiserum was used for Western blot analyses to demonstrate that a protein which is cross reactive with T7 RNA polymerase is produced. The molecular mass of this protein is 80 kDa, a size which is consistant with the nicked form of the polymerase as is often seen when expressed in E. coli. RNA polymerase assays were used to indicate that the nicked form of T7 RNA polymerase is active and capable of incorporating labeled nucleotides into transcripts in vitro. Analysis of transgenic plants did indeed show that wound-inducible activation of the T7 RNA polymerase permits the establishment of a genetic system to overexpress genes in plants using T7 RNA polymerase(Received March 20, 1997; accepted May 2, 1997)

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