• Title/Summary/Keyword: $EF1{\alpha}$

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Molecular Systematics of the Genus Megoura (Hemiptera: Aphididae) Using Mitochondrial and Nuclear DNA Sequences

  • Kim, Hyojoong;Lee, Seunghwan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.510-522
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    • 2008
  • To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.

Ethyl acetate fraction of GGEx18 modulates fatty acid β-oxidizing enzymes (In vitro 동물세포에서 GGEx18의 ethyl acetate 분획물에 의한 지방산 β-산화효소 유전자 발현의 조절)

  • Joo, Byung-Soo;Lee, Hee-Young;Lee, Hye-Rim;Yoon, Mi-Chung;Seo, Bu-Il;Kim, Beom-Hoi;Shin, Soon-Shik
    • The Korea Journal of Herbology
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    • v.27 no.2
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    • pp.53-59
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    • 2012
  • Objectives : This study was undertaken to investigate the effects of the GGEx18 ethyl acetate fraction (EF) on lipid accumulation and gene expression of fatty acid-oxidizing enzymes using 3T3-L1 adipocytes, C2C12 skeletal muscle cells, and NMu2Li liver cells. Methods : PPAR${\alpha}$, AMPK and UCPs transactivation was examined in NMu2Li hepatocytes, C2C12 myocytes, and 3T3-L1 preadipocytes using transient transfection assays. Results : 1. Compared with control, EF significantly increased the mRNA expression of VLCAD in 3T3-L1 adipocytes. 2. Compared with control, EF (0.1 ${\mu}g/ml$) significantly inhibited lipid accumulation in 3T3-L1 adipocytes. 3. EF significantly increased the mRNA expression of AMPK${\alpha}$1, AMPK${\alpha}$2 and PPAR${\alpha}$ in C2C12 skeletal muscle cells compared with control. 4. EF significantly increased the mRNA expression of genes involved in fatty acid ${\beta}$-oxidation, such as thiolase, MCAD, and CPT-1 in C2C12 skeletal muscle cells compared with control. 5. EF significantly increased the mRNA expression of UCP2 involved in energy expenditure in C2C12 skeletal muscle cells compared with control. 6. Compared with control, EF (10 ${\mu}g/ml$) significantly inhibited lipid accumulation in C2C12 skeletal muscle cells. 7. EF (10 ${\mu}g/ml$) significantly increased the mRNA expression of ACOX, HD, VLCAD and MCAD in NMu2Li liver cells compared with control. Conclusions : These results suggest that EF may prevent obesity by increasing the mRNA expression of mitochondrial fatty acid ${\beta}$-oxidizing enzymes in 3T3-L1 adipocytes, by not only regulating the fatty acid oxidation through activation of AMPK and PPAR${\alpha}$, but also increasing the UCP2 mRNA expression in C2C12 skeletal muscle cells, and by stimulating the mRNA expression of fatty acid-oxidizing enzymes in NMu2Li liver cells.

Translation elongation factor-1A1 (eEF1A1) localizes to the spine by domain III

  • Cho, Sun-Jung;Lee, Hyun-Sook;Dutta, Samikshan;Seog, Dae-Hyun;Moon, Il-Soo
    • BMB Reports
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    • v.45 no.4
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    • pp.227-232
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    • 2012
  • In vertebrates, there are two variants of eukaryotic peptide elongation factor 1A (eEF1A; formerly eEF-$1{\alpha}$), eEF1A1 and eEF1A2, which have three well-conserved domains ($D_I$, $D_{II}$, and $D_{III}$). In neurons, eEF1A1 is the embryonic type, which is expressed during embryonic development as well as the first two postnatal weeks. In the present study, EGFP-tagged eEF1A1 truncates were expressed in cortical neurons isolated from rat embryo (E18-19). Live cell images of transfected neurons showed that $D_{III}$-containing EGFP-fusion proteins (EGFP-$D_{III}$, -$D_{II-III}$, -$D_{I-III}$) formed clusters that were confined within somatodendritic domains, while $D_{III}$-missing ones (EGFP-$D_I$, -$D_{II}$, -$D_{I-II}$) and control EGFP were homogeneously dispersed throughout the neuron including axons. In dendrites, EGFP-$D_{III}$ was targeted to the heads of spine- and filopodia-like protrusions, where it was colocalized with $SynGAP{\alpha}$, a postsynaptic marker. Our data indicate that $D_{III}$ of eEF1A1 mediates formation of clusters and localization to spines.

A Strong Transcription Activity of the Bombyx mori Elongation Factor 1α Promoter

  • Goo, Tae-Won;Kim, Sung-Wan;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Choi, Kwang-Ho;Yun, Eun-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.24 no.2
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    • pp.49-55
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    • 2012
  • We previously isolated 9 clones that show stronger signal compared to B. mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid homology with elongation factor ${\alpha}$ gene of B. mori. This clone, named $bEF1{\alpha}$ (B. mori elongation factor ${\alpha}$) was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-702/+38) in the 5'-flanking region of $bEF1{\alpha}$ gene, which has about 20 fold more intensive promoter activity than BmA3 promoter. Moreover, the $bEF1{\alpha}$ promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that $bEF1{\alpha}$ promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.

A Study of Titanium Phase Transition through In-situ EF-TEM Heating Experiments (EF-TEM 직접가열 실험을 통한 titanium의 고온 상전이 연구)

  • Kim, Jin-Gyu;Lee, Young-Bu;Kim, Youn-Joong
    • Applied Microscopy
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    • v.33 no.1
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    • pp.49-58
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    • 2003
  • The ${\alpha}-{\beta}$ phase transition of titanium was investigated through in-situ EF-TEM heating experiments. Three different areas of a titanium foil were observed to minimize statistical errors. Systematic recording of diffraction patterns and images was carried out from $RT{\rightarrow}600^{\circ}C{\rightarrow}900^{\circ}C{\rightarrow}RT$ on each area. The following results were obtained: (1) Transition of titanium takes place very rapidly at $900^{\circ}C$. Two phases of titanium, ${\alpha}\;and\;{\beta}$, coexist at this temperature. (2) The transited ${\beta}$-phase appears in the form of twinned plates which are arranged in rotation relationship one another. (3) Analyses of electron diffraction patterns and EDS data indicate that the thermal oxidation layer is gradually formed on the surface of titanium above $900^{\circ}C$, which hinders the reversible ${\beta}{\rightarrow}{\alpha}$ phase transition upon cooling.

Endogenous Proteinaceous Inhibitor for Protein Methylation Reactions

  • Paik, Woon-Ki;Lee, Hyang-Woo;Kim, Sangduk
    • Archives of Pharmacal Research
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    • v.10 no.3
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    • pp.193-196
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    • 1987
  • Protein methylation occurs ubiquitously in nature and involves N-methylation of lysine, arginine, histidine, alanine, proline and glutamine, O-methylesterfication o dicarboxylic acids, and S-methylation of cysteine and methionine. In nature, methylated amino acids accur in highly specialized proteins such as histones, flagella proteins, myosin, actin, ribosomal proteins. hn RNA-bound protein, HMG-1 and HMG-2 protein, opsin, EF-Tu, EF-$1\alpha$, porcine heart citrate synthase, calmodulin, ferredoxin, $1\alpha$-amylase, heat shock protein, scleroderma antigen, nucleolar protein C23 and IF-3l.

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Comparison of RAPD, AFLP, and EF -1 α Sequences for the Phylogenetic Analysis of Fusarium oxysporum and Its formae speciales in Korea

  • Park, Jae-Min;Kim, Gi-Young;Lee, Song-Jin;Kim, Mun-Ok;Huh, Man-Kyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.34 no.2
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    • pp.45-55
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    • 2006
  • Although Fursarium oxysporum causes diseases in economically important plant hosts, identification of F. oxysporum formae speciales has been difficult due to confusing phenotypic classification systems. To resolve these complexity, we evaluated genetic relationship of nine formae speciales of F. oxysporum with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and translation elongation factor-l alpha ($EF-1{\alpha}$) gene. In addition, the correlation between mycotoxin content of fusaric acid and isolates based on molecular marker data was evaluated using the modified Mantel's test. According to these result, these fusaric acid-producing strains could not identify clearly, and independent of geographic locations and host specificities. However, in the identification of F. oxysporum formae speciales, especially, AFLP analysis showed a higher discriminatory power than that of a the RAPD and $EF-1{\alpha}$ analyses, all three techniques were able to detect genetic variability among F. oxysporum formae speciales in this study.

Phylogenetic analysiccccccccc of the genus Stemphylium based on elongation factor -1 alpha and calmodulin gene squences

  • Kong, D.W.;Cho, H.S.;Yu, S.H.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.117.2-117
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    • 2003
  • The importance and diversity of the genus Stemphylium highlights the need for accurate identification of species. However, many Stemphylium isolates have been misidentified due to the use of spore size as the only identifying character. Molecular phylogenetic analyses were performed on fifty-four isolates covering 9 Stemphylium species collected in Korea. Phylogenetic analysis of the translation elongation factor -1 alpha (EF-1) and the calmodulin gene sequence data showed that Stemphylium species were segregated into seven distinct groups, most of w hichcorrelated with species identified by morphology. Analysis of EF-1 in particular was useful for establishing well- supported relationships among the species of Stemphylium.

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Generation of Female Porcine Fibroblasts Expressing Efficiently Membrane Cofactor Protein at ${\alpha}1$,3-Galactosyltransferase locus (${\alpha}1$,3-Galactosyltransferase 유전자 좌위에서 Membrane Cofactor Protein을 효과적으로 발현하는 자성 돼지 섬유아세포의 생산)

  • Oh, Keon Bong;Kim, Bella;Hwang, Seongsoo;Ock, Sun-A;Im, Seoki;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.289-295
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    • 2013
  • Xenotransplantation of pig organs into primates results in fatal damage, referred as hyperacute rejection (HAR), and acute humoral xenograft rejection (AHXR), to the organ graft mediated by antibodies pre-existing and newly-producing in primates against their cognate pig antigens. Functional ablation of ${\alpha}1$,3-galactosyltransferase (Gal-T KO) of pig which is an enzyme involved in synthesis of Gala1-3Galb1-4GlcNAc-R antigen is essentially required to prevent HAR. Moreover, additional genetic modification under Gal-T KO background for enforced expression of human complement regulatory proteins which can inhibits complement activation is known to effectively imped HAR and AHXR. In this study, we constructed a membrane cofactor protein (MCP) expression cassette under control of human $EF1{\alpha}$ promoter. This cassette was inserted between homologous recombination regions corresponding to Gal-T locus. Subsequently this vector was introduced into ear skin fibroblasts of female pig by nucleofection. We were able to obtained 40 clones by neomycin selection and 4 clones among them were identified as clones targeted into Gal-T locus of MCP expression cassette by long-range PCR. Real time RT-PCR was shown to down-regulation of Gal-T expression. From these results, we demonstrated human $EF1{\alpha}$ promoter could induce efficient expression of MCP on cell surface of fibroblasts of female pig.

Evaluation of Evodiae Fructus Extract on the Chronic Acid Reflux Esophagitis in Rats (오수유(吳茱萸) 추출물이 만성 역류성 식도염 흰쥐에 미치는 효능 평가)

  • Lee, Jin A;Park, Hae-Jin;Kim, Soo Hyun;Kim, Min Ju;Kim, Kyeong Jo;Shin, Mi-Rae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.34 no.2
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    • pp.15-23
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    • 2019
  • Objective : Reflux esophagitis (RE) is a disease that caused gastric acid reflux and inflammation due to unstable gastroesophageal sphincter, as increasing worldwide respectively. This study was conducted to evaluate the effect of Evodiae Fructus (EF) extract on chronic reflux esophagitis in rats. Methods : The EF was measured antioxidant activity, such as total polyphenol and total flavonoid contents, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-enzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity. Rats were divided into 3 groups; Nor (normal group), Con (chronic acid reflux esophagitis rats treatment with water), EF (chronic acid reflux esophagitis rat treatment with EF 200 mg/kg body weight group). A surgically-induced chronic acid reflux esophagitis (CARE) model was established in SD rats, and treated with water or EF 200 mg/kg body weight for 14 consecutive days. Results : Administration of EF to rats of induction of chronic acid reflux esophagitis was found to reduce esophagus tissues injury. Reactive oxygen species (ROS) and produces peroxynitrite ($ONOO^-$) levels of esophagus tissues were significantly decreased in EF compared to Con group. As results of esophagus protein analyses, EF effectively reduce inflammatory-related factors ($NF-{\kappa}Bp65$, $p-I{\kappa}B{\alpha}$, iNOS, $TNF-{\alpha}$, IL-6), and increase anti-oxidant enzyme (Nrf2, HO-1, SOD, catalase, GPx-1/2). Conclusions : These results suggest that EF administration comfirmed that decreased esophagus tissues injury, oxidantive stress, anti-inflammation effect, and increased anti-oxidant effect. Therefore, EF was the potential to be used as a natural therapeutic drug.