• Molecules and Cells
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• v.23 no.1
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• pp.17-22
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• 2007

We have expressed human erythropoietin (EPO) in transgenic mice using a recombinant EPO cDNA combined with a partial TPO construct. The gene was microinjected using standard techniques and five mice were detected as transgenic by PCR and further used as founders. The life span of the transgenic founders was much shorter than that of their normal littermates. Most of the tissues of the transgenic founders contained human EPO transcripts as judged by RT-PCR. Especially high expression levels were seen in the liver and lung. EPO protein levels in serum were examined by ELISA and ranged from 266-414 mIU/ml. The number of red blood cell, white blood cell and hemoglobin in the hEPO transgenic mice was higher than in normal mice. These results indicate that overexpression of hEPO is deleterious and can provoke lung failure and erythrocytosis.

• Journal of Educational Research in Mathematics
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• v.11 no.1
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• pp.11-35
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• 2001

Many teachers report familiarity with and adherence to reform ideas, but their actual teaching practices do not reflect a deep understanding of reform. Given the challenges in implementing reform, this study intended to explore the breakdown that may occur between teachers' adoption of reform objectives and their successful incorporation of reform ideals. To this end, this study compared and contrasted the classroom social norms and sociomathematical norms of two United States second-grade teachers who aspired to implement reform. This study is an exploratory, qualitative, comparative case study. This study uses the grounded theory methodology based on the constant comparative analysis for which the primary data sources were classroom video recordings and transcripts. The two classrooms established similar social norms including an open and permissive learning environment, stressing group cooperation, employing enjoyable activity formats for students, and orchestrating individual or small group session followed by whole group discussion. Despite these similar social participation structures, the two classes were remarkably different in terms of sociomathematical norms. In one class, the students were involved in mathematical processes by which being accurate or automatic was evaluated as a more important contribution to the classroom community than being insightful or creative. In the other class, the students were continually engaged in significant mathematical processes by which they could develop an appreciation of characteristically mathematical ways of thinking, communi-eating, arguing, proving, and valuing. It was apparent from this study that sociomathematical norms are an important construct reflecting the quality of students' mathematical engagement and anticipating their conceptual learning opportunities. A re-theorization of sociomathematical norms was offered so as to highlight the importance of this construct in the analysis of reform-oriented classrooms.

• The Plant Pathology Journal
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• v.23 no.2
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• pp.76-89
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• 2007

CaCDPK4, a full-length cDNA clone encoding Capsicum annuum calcium-dependent protein kinase 4, was isolated from chili pepper (Capsicum annuum L.). Deduced amino acid sequence of CaCDPK4 shares the highest homology with tobacco NpCDPK8 and chickpea CaCDPK2 with 79% identity. Genomic blot analyses revealed that CaCDPK4 is present as a single copy in pepper genome, but it belongs to a multigene family. CaCDPK4 was highly induced when pepper plants were inoculated with an incompatible bacterial pathogen. Induced levels of CaCDPK4 transcripts were also detected in pepper leaves by the treatment of ethephon, an ethylene-inducing agent, and high-salt stress condition. The bacterial-expressed GST-CaCDPK4 protein showed to retain the autophosphorylation activity in vitro. GUS expression driven by CaCDPK4 promoter was examined in transgenic Arabidopsis containing transcriptional fusion of CaCDPK4 promoter. GUS expression under CaCDPK4 promoter was strong in the root and veins of the seedlings. GW (-1965) and D3 (-1377) promoters conferred on GUS expression in response to inoculation of an incompatible bacterial pathogen, but D4-GUS (-913) and DS-GUS (-833) did not. Taken together, our results suggest that CaCDPK4 can be implicated on signal transduction pathway of defense response against an incompatible bacterial pathogen in pepper.

• The Plant Pathology Journal
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• v.33 no.5
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• pp.458-466
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• 2017

Xanthomonas oryzae pv. oryzae (Xoo), the causative agent of bacterial blight, is a major threat to rice productivity. Here, we performed RNA-Seq based transcriptomic analysis of Xoo transcripts isolated under in planta growth (on both susceptible and resistant hosts) and in vitro culture conditions. Our in planta extraction method resulted in successful enrichment of Xoo cells and provided RNA samples of high quality. A total of 4,619 differentially expressed genes were identified between in planta and in vitro growth conditions. The majority of the differentially expressed genes identified under in planta growth conditions were related to the nutrient transport, protease activity, stress tolerance, and pathogenicity. Among them, over 1,300 differentially expressed genes were determined to be secretory, including 184 putative type III effectors that may be involved in Xoo pathogenicity. Expression pattern of some of these identified genes were further validated by semi-quantitative RT-PCR. Taken together, these results provide a transcriptome overview of Xoo under in planta and in vitro growth conditions with a focus on its pathogenic processes, deepening our understanding of the behavior and pathogenicity of Xoo.

• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.259-268
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• 2000

Senescence is a sequence of biochemical and physiological events that lead to death of a cell, organ, or whole organism. Senescence is now clearly regarded as a genetically determined and evolutionarilly acquired developmental process comprising the final stage of development. However, in spite of the biological and practical importance, genetic mechanism of senescence has been very limited. Through forward and reverse genetic approaches, we are trying to reveal the molecular and genetic mechanism of senescence in plants, employing leaf organs of Arabidopsis as a model system. Using forward genetic approach, we have initially isolated several delayed senescence mutants either from T-DNA insertional lines or chemical-mutagenized lines. In the case of ore 4 and ore 9 mutants, the mutated genes were identified. The recent progress on characterization of mutants and identification of the mutated genes will be reported. We are also screening mutations from other various sources of mutant pools, such as activation tagging lines and promoter trap lines. Two dominant senescence-delayed mutants were isolated from the activation tagging pool. Cloning of the genes responsible for this phenotype is in progress. For reverse genetic approach, the genes that induced during leaf senescence were first isolated by differential screening method. We are currently using PCR-based suppression subtractive hybridization, designed to enrich a cDNA library for rare differentially expressed transcripts. Using this method, we have identified over 35 new sequences that are upregulated at leaf senescence stage. We are investigating the function of these novel genes by systemically generating antisense lines.

• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.83-87
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• 2000

Carnation petals exhibit autocatalytic ethylene production and wilting during senescence. The autocatalytic ethylene production is induced by the expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase genes, whereas the wilting of petals is related to expression of the cysteine proteinase (CP) gene. Until recently, it has been believed that these two phenomena, autocatalytic ethylene production and wilting, are regulated in concert in senescing carnation petals, since the two phenomena occurred closely in parallel. Our studies with petals of a transgenic carnation harboring a sense ACC oxidase transgene and petals of carnation flowers treated with 1,1-dimethyl-4-(phenylsulfonyl) semicarbazide showed that the expression of ACC synthase and ACC oxidase genes and that of CP are regulated differently in carnation psanetals. Interestingly, in the petals of transgenic carnation, the transcript for CP was accumulated but the transcripts for ACC synthase and ACC oxidase were not accumulated in response to exogenous ethylene. Based on these results, we hypothesized that two ethylene signaling pathways, one leading to the expression of ACC synthase and ACC oxidase genes and the other leading to the expression of CP gene, are functioning in senescing carnation petals.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.149-155
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• 2010

Expression of the genes for carotenoid bio-synthesis (crt) is dependent on light, but little is known about the underlying mechanism of light sensing and signalling in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter, Synechocystis). In the present study, we investigated the light-induced increase in the transcript levels of Synechocystis crt genes, including phytoene synthase (crtB), phytoene desaturase (crtP), ${\zeta}$-carotene desaturase (crtQ), and ${\beta}$-carotene hydroxylase (crtR), during a darkto-light transition period. During the dark-to-light shift, the increase in the crt transcript levels was not affected by mutations in cyanobacterial photoreceptors, such as phytochromes (cph1, cph2 and cph3) and a cryptochrome-type photoreceptor (ccry), or respiratory electron transport components NDH and Cyd/CtaI. However, treatment with photosynthetic electron transport inhibitors significantly diminished the accumulation of crt gene transcripts. Therefore, the light induction of the Synechocystis crt gene expression is most likely mediated by photosynthetic electron transport rather than by cyanobacterial photoreceptors during the dark-to-light transition.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.462-471
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• 2017

Thiamine pyrophosphate (TPP), the active form of thiamine is a cofactor for enzymes involved in central metabolism pathways. However, it is also known to have a role as a stress signaling molecule in response to environmental changes. Anabaena sp. and N. oculata are microorganisms which are abundantly found in Malaysia's freshwater and marine ecosystem. However, not much studies have been done especially in regards to thiamine biosynthesis. This work aimed to amplify of gene transcripts coding for thiamine biosynthesis enzymes besides looking at the expression of thiamine biosynthesis genes upon stress application. Various stress inducers were applied to the cultures and RNA was extracted at different time points. The first two genes, ThiC and ThiG/Thi4 encoding enzymes of the pyrimidine and thiazole branch respectively in the thiamine biosynthesis pathway were identified and amplified. The expression of the genes were analysed via RT-PCR and the intensity of bands were analysed using ImageJ software. The results showed up to 4-fold increase in the expression of ThiC and ThiG gene transcript as compared to control sample in Anabaena sp. ThiC gene in N. oculata showed an expression of 6-fold higher as compared to control sample. In conclusion, stresses induced the expression of the gene coding for one of the most important enzymes in thiamine biosynthesis pathway. This is an agreement with the hypothesis that overexpression of thiamine is crucial in assisting plants to combat abiotic stresses.

• International Journal of Industrial Entomology and Biomaterials
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• v.13 no.1
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• pp.23-30
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• 2006

BmCu/Zn SOD was isolated from early embryo of Bombyx mori using microarray analysis. The BmCu/Zn SOD gene was observed during the early embryonic stage with the strongest signal found at the unfertilizaion, fertilization and blastoderm stages. The BmCu/Zn SOD gene encodes a protein of 154 amino acids with a calculated Mr of 15 kDa. The deduced amino acid sequence of BmCu/Zn SOD indicated that the residues that form on the Cu/Zn binding site are conserved and that the sequence is a 60% identity to that of M. domestica. In a phylogenetic tree, Bm SOD was also close to Drosophila SODs rather than other insect SODs. The BmCu/Zn SOD gene exists as a single copy in the genome. Transcripts of BmCu/Zn SOD cDNA were identified by northern blot analysis. The expression of the BmCu/Zn SOD gene was observed weakly in most of larvae, pre-pupae, pupae and adult tissues. Also, the BmCu/Zn SOD gene was observed in early embryonic stage. Although the roles of SODs remains to be further elucidated, the high expression of BmCu/Zn SOD gene at before 24 h post fertilization suggests that this gene is of general importance during early embryogenesis in the Bombyx mod.

• BMB Reports
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• v.41 no.3
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• pp.259-265
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• 2008

Three peroxidase (POD) cDNAs were isolated from dehydration-treated fibrous roots of sweetpotato (Ipomoea batatas) plant via the screening of a cDNA library, and their expressions were assessed to characterize functions of each POD in relation to environmental stress. Three PODs were divided into two groups, designated the basic PODs (swpb4, swpb5) and the anionic PODs (swpa7), on the basis of the pI values of mature proteins. Fluorescence microscope analysis indicated that three PODs are secreted into the extracellular space. RT-PCR analysis revealed that POD genes have diverse expression patterns in a variety of plant tissues. Swpb4 was abundantly expressed in stem tissues, whereas the expression levels of swpb5 and swpa7 transcripts were high in fibrous and thick pigmented roots. Swpb4 and swpa7 showed abundant expression levels in suspension cultured cells. Three POD genes responded differently in the leaf and fibrous roots in response to a variety of stresses including dehydration, temperature stress, stress-associated chemicals, and pathogenic bacteria.