• Title/Summary/Keyword: Resistance marker

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Development of Solanum hougasii-specific markers using the complete chloroplast genome sequences of Solanum species (엽록체 전장유전체 정보를 이용한 Solanum hougasii 특이적 분자마커 개발)

  • Kim, Soojung;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.141-149
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    • 2020
  • Solanum hougasii, one of the wild Solanum species, has been widely used in potato breeding since it exhibits excellent resistance to diverse important pathogens. S. hougasii can be directly crossed with the cultivated tetraploid potato (S. tuberosum) owing to its EBN (Endosperm Balanced Number) value of 4, which is same as that of S. tuberosum although it is an allohexaploid. In this study, the complete chloroplast genome sequence of S. hougasii was obtained by next-generation sequencing technology, and compared with that of the chloroplast genome of seven other Solanum species to identify S. hougasii-specific PCR markers. The length of the complete chloroplast genome of S. hougasii was 155,549 bp. The structural organization of the chloroplast genome in S. hougasii was found to be similar to that of seven other Solanum species studied. Phylogenetic analysis of S. hougasii with ten other Solanaceae family members revealed that S. hougasii was most closely related to S. stoloniferum, followed by S. berthaultii, and S. tuberosum. Additional comparison of the chloroplast genome sequence with that of five other Solanum species revealed five InDels and 43 SNPs specific to S. hougasii. Based on these SNPs, four PCR-based markers were developed for the differentiation of S. hougasii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary and breeding aspects of Solanum species.

Endophytic bacterium Pseudomonas fluorescens strain EP103 was effective against Phytophthora capsici causing blight in chili pepper (식물근권에서 분리한 Pseudomonas fluorescens strain EP103에 의한 고추역병억제)

  • Kim, Tack-Soo;Dutta, Swarnalee;Lee, Se Won;Park, Kyungseok
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.422-428
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    • 2014
  • Endophytic bacterial strains from root tissue of strawberry were screened for their efficacy in growth improvement and control of Phytophthora blight disease of chili pepper plant under greenhouse condition. Plants treated with the strain EP103, identified as Pseudomonas fluorescens, showed growth improvement in terms of fresh weight and root length compared to the untreated control and other endophytic strains. When challenged with Phytophthora capsici, there was significant reduction of disease in EP103 treated plants with an efficacy of 78.7%. There was no direct inhibition of the target pathogen by EP103 when tested under in vitro antibiosis assay. Analysis of differential expression of selected marker genes for induced systemic resistance (ISR) in plants treated with EP103 and challenged with P. capsici showed up-regulation of PR1 and PR10 pathogenesis-related (PR) proteins. PCR analysis showed that EP103 produced secondary metabolites such as pyoluteorin, pyrrolnitrin, hydrogen cyanide and orfamide A. This study indicated the potential of endophytic P. fluorescens strain EP103 as an efficient biocontrol agent against P. capsici in chili pepper plant.

Development of an In Vitro Test System Measuring Transcriptional Downregulatory Activities on IL-13

  • Choi, Jeong-June;Park, Bo-Kyung;Park, Sun-Young;Yun, Chi-Young;Kim, Dong-Hee;Kim, Jin-Sook;Hwang, Eun-Sook;Jin, Mi-Rim
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.331-337
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    • 2009
  • Interleukin-13 (IL-13) has been proposed as a therapeutic target for bronchial asthma as it plays crucial roles in the pathogenesis of the disease. We developed an in vitro test system measuring transcriptional downregulatory activities on IL-13 as a primary screening method to select drug candidates from natural products. The promoter region of IL-13 (-2,048 to +1) was cloned into the upstream of a luciferase gene in the plasmid pGL4.14 containing the hygromycin resistance gene as a selection marker, generating pGL4.14-IL-13. The EL-4 thymoma and RBL-2H3 mast cells transiently expressing this plasmid highly produced the luciferase activities by responding to PI (PMA and ionomycin) stimulation up to 8-fold and 13-fold compared with the control, respectively, whereas cyclosporin A, a well-known antiasthmatic agent, significantly downregulated the activities. The BF1 clone of RBL-2H3 cells constitutively expressing pGL4.14-IL-13 was established by selecting surviving cells under a constant lethal dose of hygromycin treatment. The feasibility of this system was evaluated by measuring the downregulatory activities of 354 natural products on the IL-13 promoter using the BF1 clone. An extract from Morus bombycis (named TBRC 156) significantly inhibited PI-induced luciferase activities and IL-13 mRNA expression, but not the protein expression. Fisetin (named TBRC 353) inhibited not only PI-induced luciferase activities and mRNA expression, but also the IL-13 protein secretion, whereas myricetin (named TBRC 354) could not suppress the IL-13 expression at all. Our data indicated that this in vitro test system is able to discriminate the effects on IL-13 expression, and furthermore, that it might be suitable as a simple and time-saving primary screening system to select antiasthmatic agents by measuring transcriptional activities of the IL-13 promoter.

Pro-Apoptotic Role of the Human YPEL5 Gene Identified by Functional Complementation of a Yeast moh1Δ Mutation

  • Lee, Ji Young;Jun, Do Youn;Park, Ju Eun;Kwon, Gi Hyun;Kim, Jong-Sik;Kim, Young Ho
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.633-643
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    • 2017
  • To examine the pro-apoptotic role of the human ortholog (YPEL5) of the Drosophila Yippee protein, the cell viability of Saccharomyces cerevisiae mutant strain with deleted MOH1, the yeast ortholog, was compared with that of the wild-type (WT)-MOH1 strain after exposure to different apoptogenic stimulants, including UV irradiation, methyl methanesulfonate (MMS), camptothecin (CPT), heat shock, and hyperosmotic shock. The $moh1{\Delta}$ mutant exhibited enhanced cell viability compared with the WT-MOH1 strain when treated with lethal UV irradiation, 1.8 mM MMS, $100{\mu}M$ CPT, heat shock at $50^{\circ}C$, or 1.2 M KCl. At the same time, the level of Moh1 protein was commonly up-regulated in the WT-MOH1 strain as was that of Ynk1 protein, which is known as a marker for DNA damage. Although the enhanced UV resistance of the $moh1{\Delta}$ mutant largely disappeared following transformation with the yeast MOH1 gene or one of the human YPEL1-YPEL5 genes, the transformant bearing pYES2-YPEL5 was more sensitive to lethal UV irradiation and its UV sensitivity was similar to that of the WT-MOH1 strain. Under these conditions, the UV irradiation-induced apoptotic events, such as FITC-Annexin V stainability, mitochondrial membrane potential (${\Delta}{\psi}m$) loss, and metacaspase activation, occurred to a much lesser extent in the $moh1{\Delta}$ mutant compared with the WT-MOH1 strain and the mutant strain bearing pYES2-MOH1 or pYES2-YPEL5. These results demonstrate the functional conservation between yeast Moh1 and human YPEL5, and their involvement in mitochondria-dependent apoptosis induced by DNA damage.

Treatment of Metastatic Colorectal Cancer With or Without Bevacizumab: Can the Neutrophil/Lymphocyte Ratio Predict the Efficiency of Bevacizumab?

  • Dirican, Ahmet;Varol, Umut;Kucukzeybek, Yuksel;Alacacioglu, Ahmet;Erten, Cigdem;Somali, Isil;Can, Alper;Demir, Lutfiye;Bayoglu, Ibrahim Vedat;Akyol, Murat;Yildiz, Yasar;Koyuncu, Betul;Coban, Eyup;Tarhan, Mustafa Oktay
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4781-4786
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    • 2014
  • Background: The purpose of this study was to analyze the predictive value of neutrophil/lymphocyte ratio (NLR) to better clarify which patient groups will benefit the most from particular treatments like bevacizumab. Materials and Methods: A total of 245 treatment-naive metastatic colorectal cancern (mCRC) patients were retrospectively enrolled and divided into 2 groups: 145 group A patients were treated with chemotherapy in combination with bevacizumab, and 100 group B patients were treated as above without bevacizumab. Results: Group A patients had better median overall survival (OS) and progression-free survival (PFS) (24.0 and 9.0 months) than group B patients (20 and 6.0 months) (p=0.033; p=0.015). In patients with low NLR, OS and PFS were significantly longer in group A patients (27 vs 18 months, p=0.001; 11 vs 7 months, p=0.017). Conclusions: We conclude that NLR, a basal cancer related inflammation marker, is associated with the resistance to bevacizumab-based treatments in mCRC patients.

Effects of Head-down Tilt $(-6^{\circ})$ on Peripheral Blood Flow in Dogs (두부하위$(-6^{\circ})$로의 체위변동이 말초혈류에 미치는 영향)

  • Chae, E-Up;Yang, Seon-Young;Bae, Jae-Hoon;Song, Dae-Kyu
    • The Korean Journal of Physiology
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    • v.24 no.1
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    • pp.51-65
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    • 1990
  • The purpose of the present study was to examine the hemodynamic responses, especially in arterial and skin blood flows, in conjunction with the changes of plasma catecholamine levels as an indirect marker of adrenergic tone during the early stage of head-down tilt (HDT), and to evaluate the early physiological regulatory mechanism in simulated weightlessness. Ten mongrel dogs, weighing8\;{\sim}\;14\;kg, were intravenously anesthetized with nembutal, and postural changes were performed by using the tilting table. The postural changes were performed in the following order: supine, prone, HDT $(-6^{\circ}C)$ and lastly recovery prone position. The duration of each position was 30 minutes. The measurements were made before, during and after each postural change. The arterial blood flow $({\.{Q}})$ at the left common carotid and right brachial arteries was measured by the electromagnetic flowmeter. Blood pressure (BP) was directly measured by pressure transducer in the left brachial artery. To evaluate the peripheral blood flow, skin blood flow $({\.{Q}})$ was calculated by the percent changes of photoelectric pulse amplitude on the forepaw, and skin temperature was recorded. The peripheral vascular resistance (PR) was calculated by dividing respective mean BP values by ${\.{Q}}$ of both sides of common carotid and brachial arteries. Heart rate (HR), respiratory rate (f) and PH, $Po_{2},\;Pco_{2}$ and hematocrit of arterial and venous blood were also measured. The concentration of plasma epinephrine and norepinephrine was measured by radioenzymatic method. The results are summarized as follows: Tilting to head-down position from prone position, HR was initially increased (p<0.05) and BP was not significantly changed. While ${\.{Q}}$ of the common carotid artery was decreased (p<0.05) and PR through the head was increased, ${\.{Q}}$ of the brachial artery was increased (p<0.05) and PR through forelimbs was decreased. ${\.{Q}}$ of the forepaw was initially increased (p<0.05) and then slightly decreased, on the whole revealing an increasing trend. Plasma norepinephrine was slightly decreased and the epinephrine was slightly increased. f was increased and arterial pH was increased (p<0.05). In conclusion, the central blood pooling during HDT shows an increased HR via Bainbridge reflex and an increased ${\.{Q}}$ of the forepaw and brachial ${\.{Q}}$, due to decreased PR which may be originated from the depressor reflex of cardiopulmonary baroreceptors. It is suggested that the blood flow to the brain was adequately regulated throughout HDT $(-6^{\circ}C)$ in spite of central blood pooling. And it is apparent that the changes of plasma norepinephrine level are inversely proportional to those of ${\.{Q}}$ of the forepaw, and the changes of epinephrine level are paralleled with those of the brachial ${\.{Q}}$.

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Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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Field Performance and Morphological Characterization of Transgenic Codonopsis lanceolata Expressing $\gamma-TMT$ Gene.

  • Ghimire, Bimal Kumar;Li, Cheng Hao;Kil, Hyun-Young;Kim, Na-Young;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Chung, Ill-Min;Lee, Sun-Joo;Eom, Seok-Hyun;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.339-345
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    • 2007
  • Field performance and morphological characterization was conducted on seven transgenic lines of Codonopsis lanceolata expressing ${\gamma}-TMT$ gene. The shoots were obtained from leaf explants after co-cultivation with Agrobacterium tume-faciens strain LBA 4404 harboring a binary vector pYBI 121 that carried genes encoding ${\gamma}-Tocopherol$ methyltransferase gene (${\gamma}-TMT$) and a neomycin phosphotransferase II gene (npt II) for kanamycin resistance. The transgenic plants were transferred to a green house for acclimation. Integration of T-DNA into the $T_0\;and\;T_1$ generation of transgenic Codonopsis lanceolata genome was confirmed by the polymerase chain reaction and southern blot analysis. The progenies of transgenic plants showed phenotypic differences within the different lines and with relative to control plants. When grown in field, the transgenic plants in general exhibited increased fertility, significant improvement in the shoot weight, root weight, shoot height and rachis length with relation to the control plants. However, all seven independently derived transgenic lines produced normal flower with respect to its shape, size, color and seeds number at its maturity. Indicating that the addition of a selectable marker gene in the plant genome does not effect on seed germination and agronomic performance of transgenic Codonopsis lanceolata. $T_1$ progenies of these plants were obtained and evaluated together with control plant in a field experiment. Overall, the agronomic performance of $T_1$ progenies of transgenic Codonopsis lanceolata showed superior to that of the seed derived non-transgenic plant. In this study, we report on the morphological variation and agronomic performance of transgenic Codonopsis lanceolata developed by Agrobacterium transformation.

Deubiquitinase Otubain 1 as a Cancer Therapeutic Target (암 치료 표적으로써 OTUB1)

  • Kim, Dong Eun;Woo, Seon Min;Kwon, Taeg Kyu
    • Journal of Life Science
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    • v.30 no.5
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    • pp.483-490
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    • 2020
  • The ubiquitin system uses ligases and deubiquitinases (DUBs) to regulate ubiquitin position on protein substrates and is involved in many biological processes which determine stability, activity, and interaction of the target substrate. DUBs are classified in six groups according to catalytic domain, namely ubiquitin-specific proteases (USPs); ubiquitin C-terminal hydrolases (UCHs); ovarian tumor proteases (OTUs); Machado Joseph Disease proteases (MJDs); motif interacting with Ub (MIU)-containing novel DUB family (MINDY); and Jab1/MPN/MOV34 metalloenzymes (JAMMs). Otubain 1 (OTUB1) is a DUB in the OTU family which possesses both canonical and non-canonical activity and can regulate multiple cellular signaling pathways. In this review, we describe the function of OTUB1 through regulation of its canonical and non-canonical activities in multiple specifically cancer-associated pathways. The canonical activity of OTUB1 inhibits protein ubiquitination by cleaving Lys48 linkages while its non-canonical activity prevents ubiquitin transfer onto target proteins through binding to E2-conjugating enzymes, resulting in the induction of protein deubiquitination. OTUB1 can therefore canonically and non-canonically promote tumor cell proliferation, invasion, and drug resistance through regulating FOXM1, ERα, KRAS, p53, and mTORC1. Moreover, clinical research has demonstrated that OTUB1 overexpresses with high metastasis in many tumor types including breast, ovarian, esophageal squamous, and glioma. Therefore, OTUB1 has been suggested as a diagnosis marker and potential therapeutic target for oncotherapy.

Association of Adiponectin Polymorphisms with Type 2 Diabetes in Korean Population (한국인에서의 아디포넥틴의 유전자다형성과 제2형 당뇨병과의 연관성)

  • Yoo, Min;Kim, Su-Won
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1495-1498
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    • 2009
  • Type 2-diabetes is a typical polygenic disease complex, for which several common risk alleles have been identified. Adiponectin, which modulates insulin resistance as well as glucose and lipid metabolism, has recently been associated with type 2-diabetes (T2DM). Therefore, we investigated the genotype for the T45G and G267T polymorphisms in adiponectin genes in the Korean population and compared genotypes of patients with those of a control group. 100 patients (63 male, 37 female), who previously underwent T2DM and 100 controls (36 male, 63 female) participated in this study. There was a strong association between T45G polymorphism in the adiponectin gene and T2DM. The present study shows that adiponectin T45G polymorphism may be associated with the pathogenesis of T2DM. Further studies with a larger population may be needed for the development of diagnostic methods at genetic levels such as DNA chip.