• Title/Summary/Keyword: that-Deletion

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Analysis of polymorphic region of GAM-1 gene in Plasmodium vivax Korean isolates

  • Kho, Weon-Gyu;Chung, Joon-Yong;Hwang, Ui-Wook;Chun, Jin-Ho;Park, Yeong-Hong;Chung, Woo-Chul
    • Parasites, Hosts and Diseases
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    • v.39 no.4
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    • pp.313-318
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    • 2001
  • The identification , characterization and quantification of Plasmodium sp. genetic polymorphism are becoming increasingly important in the vaccine development. We investigated polymorphism of Plasmodium vivax GAM-1 (PvGAM-1) gene in 30 Korean isolates. The polymorphic region of the PvGAM-1 gene, corresponding to nt 3792-4029, was amplified using polymerase chain reaction (PCR) followed by sequencing. All of the P. viuax Korean isolates were one type of GAM-1 gene, which were identical to that of the Belem strain. It is suggested that PvGAM-1 could not be used as a genetic marker for identifying or classifying P. vivax Korean isolates. It revealed that the polymorphic pattern as acquired basically by duplication and modification or deletion event of a 33 bp-motif fragment ended by poly guanine (G) and that there were at least three complete and one partial 33 Up-motif sequences within the polymorphic region in the longest cases such as those of South Korean and Belem isolates. In addition, we clustered P. vivax isolates with parsimonious criteria on the basis of PvGAM- 1 polymorphic patterns (insertion/deletion patterns) .

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ILLEGITIMATE RECOMBINATION MEDIATED BY MAMMALIAN DNA TOPOISOMERASE II

  • BAE YOUNG-SEUK;IKEDA HIDEO
    • Proceedings of the Microbiological Society of Korea Conference
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    • 1991.04a
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    • pp.82-96
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    • 1991
  • To understand the mechanism of illegitimate recombination in mammalian cells, we have examined the recombination role of DNA topoisomerase II (Topo II ). We found that purified calf thymus Topo II mediates recombination between two phage $\lambda$ DNA molecules in an in vitro system. The enzyme mainly produced a linear monomer recombinant DNA that can be packaged in vitro. Novobiocin and anti-calf thymus Topo II antibody inhibit this ATP-dependent recombination. The recombinant molecules contain duplications or deletion, and most crossovers take place between nonhomologous sequences of $\lambda$ DNA, as judged by the sequences of recombination junctions. In order to study the effects of Topo II on illegitimate recombination in mammalian cells, we have developed a new shuttle vector, pNKl, which contains three bacterial genes, amp(APR), galK and neo($Km^R$). Using this system, we have shown that a Topo II inhibitor, VM26, stimulated deletion formation in pNK1 DNA in monkey COS1 cells. Both in vitro and in vivo results suggest that Topo II participates in illegitimate recombination in mammalian cells.

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Identification of a Glucokinase that Generates a Major Glucose Phosphorylation Activity in the Cyanobacterium Synechocystis sp. PCC 6803

  • Lee, Jung-Mi;Ryu, Jee-Youn;Kim, Hyong-Ha;Choi, Sang-Bong;de Marsac, Nicole Tandeau;Park, Youn-Il
    • Molecules and Cells
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    • v.19 no.2
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    • pp.256-261
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    • 2005
  • In silico analysis of genome of the cyanobacterium Synechocystis sp. PCC 6803 identified two genes, slr0329 and sll0593, that might participate in glucose (Glc) phosphorylation (www.kazusa.or.jp/cyano). In order to determine the functions of these two genes, we generated deletion mutants, and analyzed their phenotypes and enzymatic activities. In the presence of 10 mM Glc, wild-type (WT) and slr0329 defective strain (M1) grew fast with increased respiratory activity and NADPH production, whereas the sll0593 deletion mutant (M2) failed to show any of the Glc responses. WT and M1 were not significantly different in their glucokinase activity, but M2 had 90% less activity. Therefore, we propose that Sll0593 plays a major role in the phosphorylation of glucose in Synechocystis cells.

The Influence of the N-Terminal Region of Antimicrobial Peptide Pleurocidin on Fungal Apoptosis

  • Choi, Hyemin;Lee, Dong Gun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1386-1394
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    • 2013
  • In our previous study, the 25-mer antimicrobial peptide pleurocidin (Ple) had been thought to induce apoptosis in Candida albicans. This study demonstrated that reactive oxygen species (ROS) production was a major cause of Ple-induced apoptosis. Four truncated analogs were synthesized to understand the functional roles in the N- and C-terminal regions of Ple on the apoptosis. Ple, Ple (4-25), Ple (1-22), and Ple (1-19) produced ROS, including hydroxyl radicals, on the order of [Ple > Ple (1-22) > Ple (4-25) > Ple (1-19)], whereas Ple (7-25) did not induce any ROS production. The results suggested that the N-terminal deletion affected the ROS-inducing activities much more than that of the C-terminal deletion, and net hydrophobicity [Ple > Ple (1-22) > Ple (4-25) > Ple (1-19) > Ple (7-25)] was related to ROS generation rather than other primary factors like net charge. Hence, we focused on the N-terminal-truncated peptides, Ple (4-25) and Ple (7-25), and examined other apoptotic features, including mitochondrial membrane depolarization, caspase activation, phosphatidylserine externalization, and DNA and nuclear fragmentation. The results also confirmed the disappearance of apoptotic activity of Ple (7-25) by the truncation of the N-terminal region (1-6) and the specific activity patterns between Ple and analogs. In conclusion, the N-terminal region of Ple played an important role in apoptosis.

Mind Bomb-2 Regulates Hippocampus-dependent Memory Formation and Synaptic Plasticity

  • Kim, Somi;Kim, TaeHyun;Lee, Hye-Ryeon;Kong, Young-Yun;Kaang, Bong-Kiun
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.6
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    • pp.515-522
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    • 2015
  • Notch signaling is a key regulator of neuronal fate during embryonic development, but its function in the adult brain is still largely unknown. Mind bomb-2 (Mib2) is an essential positive regulator of the Notch pathway, which acts in the Notch signal-sending cells. Therefore, genetic deletion of Mib2 in the mouse brain might help understand Notch signaling-mediated cell-cell interactions between neurons and their physiological function. Here we show that deletion of Mib2 in the mouse brain results in impaired hippocampal spatial memory and contextual fear memory. Accordingly, we found impaired hippocampal synaptic plasticity in Mib2 knock-out (KO) mice; however, basal synaptic transmission did not change at the Schaffer collateral-CA1 synapses. Using western blot analysis, we found that the level of cleaved Notch1 was lower in Mib2 KO mice than in wild type (WT) littermates after mild foot shock. Taken together, these data suggest that Mib2 plays a critical role in synaptic plasticity and spatial memory through the Notch signaling pathway.

p53 Mutations in Ewing's Sarcoma (유잉육종의 p53 돌연변이)

  • Bae, Dae-Kyung;Sun, Seung-Deok
    • The Journal of the Korean bone and joint tumor society
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    • v.6 no.4
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    • pp.143-151
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    • 2000
  • Purpose : The p53 tumor suppressor gene is one of the most frequently altered genes in human malignancies. We try to explore the implication of p53 alteration in Ewing's sarcoma. Materials and Methods : We analyzed 35 paraffin blocks to explore the deletion and sequence alterations of p53. Results : Quantitative PCR analysis showed that 2 tumors showed a homozygous deletion of the gene. Mutational analysis of exons 4 to 9 of p53 by PCR-SSCP revealed that 3 tumors carry sequence alterations in exons 5 or 8, and DNA sequencing analysis identified missense point mutations. Conclusion : Taken together, our data demonstrate that p53 is genetically altered in a small fraction of Ewing's sarcoma.

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Control of Singlet Oxygen-induced Oxidative Damage in Escherichia coli

  • Kim, Sun-Yee;Kim, Eun-Ju;Park, Jeen-Woo
    • BMB Reports
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    • v.35 no.4
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    • pp.353-357
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    • 2002
  • Singlet oxygen ($^1O_2$) is highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules. The oxyR gene product regulates the expression of the enzymes and proteins that are needed for cellular protection against oxidative stress. In this study, the role of oxyR in cellular defense against a singlet oxygen was investigated using Escherichia coli oxyR mutant strains. Upon exposure to methylene blue and visible light, which generates singlet oxygen, the oxyR overexpression mutant was much more resistant to singlet oxygen-mediated cellular damage when compared to the oxyR deletion mutant in regard to growth kinetics, viability and protein oxidation. Induction and inactivation of major antioxidant enzymes, such as superoxide desmutase and catalase, were observed after their exposure to a singlet oxygen generating system in both oxyR strains. However, the oxyR overexpression mutant maintained significantly higher activities of anticxidant enzymes than did the oxyR deletion mutant. These results suggest that the oxyR regulon plays an important protective role in singlet oxygen-mediated cellular damage, presumably through the protection of antioxidant enzymes.

A Study on the Characteristics for the Blanking of Lead Frame with the nickel alloy Alloy42 (니켈합금 Alloy42를 사용하는 리드프레임의 블랭킹 특성에 관한 기초연구)

  • Bahn Gab-su;Suh Eui-kwon;Lee Gwang-ho;Mo Chang-ki
    • Transactions of the Korean Society of Machine Tool Engineers
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    • v.13 no.6
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    • pp.87-93
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    • 2004
  • An experimental is carried out to investigate the characteristics of blanking for nickel alloy Alloy42 (t=0.203mm), a kind of IC lead frame material. By varying clearance between die and punch the shapes of shear profile are examined. Finite element analysis with element deletion algorithm for ductile fracture mode is also carried out to study the effect of clearance theoretically and to compare with experimental results. The rectangular shape specimen with four different comer radius is used to study the characteristics of blanking for straight side and comer region simultaneously. As the result the ratios measured k(m experiment of roll over, burnish and fracture zone based on initial blank thickness are compared with those of FE analysis. Both experiment and FE analysis show that the amount of mil over and fracture is increased as the clearance increases. When the radius of comer is less than thickness of blank it has been found that larger clearance is required than that of straight region in order to maintain same quality of shear profile at the comer region.

A Study on the Characteristics for the Blanking of Lead Frame with the Rectangular Shape Blanking (사각형 블랭킹을 통한 리드프레임의 블랭킹 특성에 관한 기초연구)

  • Lim, San-Heon;Suh, Eui-Kwon;Shim, Hyun-Bo
    • Journal of the Korean Society for Precision Engineering
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    • v.18 no.3
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    • pp.182-188
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    • 2001
  • An experiment is carried out to investigate the characteristics of blanking for copper alloy C194 (t=0.254mm), a kind of IC lead frame material. By varying clearance between die and punch, the shapes of shear profile are examined. Finite element analysis with element deletion algorithm for ductile fracture mode is also carried out to study the effect of clearance theoretically and to compare with experimental results. The rectangular shape specimen with four different corner radius is used to study the characteristics of blanking for straight side and corner region simultaneously. As the result, the ratios measured from the experiment of roll over, burnish, and fracture zone based on intial blank thickness are compared with those of FE analysis. Both experiment and FE analysis show that the amount of roll over and fracture is increased as the clearance increases. It has been found that larger clearance is required than that of straight region when the radius of corner is less than thickness of blank, in order to maintain same quality of shear profile at the corner region.

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OxyR Regulon Controls Lipid Peroxidation-mediated Oxidative Stress in Escherichia coli

  • Yoon, Seon-Joo;Park, Ji-Eun;Yang, Joon-Hyuck;Park, Jeen-Woo
    • BMB Reports
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    • v.35 no.3
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    • pp.297-301
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    • 2002
  • Membrane lipid peroxidation processes yield products that may react with DNA and proteins to cause oxidative modifications. The oxyR gene product regulates the expression of enzymes and proteins that are needed for cellular protection against oxidative stress. Upon exposure to tert-butylhydroperoxide (t-BOOH) and 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH), which induce lipid peroxidation in membranes, the Escherichia coli oxyR overexpression mutant was much more resistant to lipid peroxidation-mediated cellular damage, when compared to the oxyR deletion mutant in regard to growth kinetics, viability, and DNA damage. The deletion of the oxyR gene in E. coli also resulted in increased susceptibility of superoxide dismutase to lipid peroxidation-mediated inactivation. The results indicate that the peroxidation of lipid is probably one of the important intermediary events in free radical-induced cellular damage. Also, the oxyR regulon plays an important protective role in lipid peroxidation-mediated cellular damage.