• Title/Summary/Keyword: B-tubulin

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Molecular Analysis of Botrytis cinerea Causing Ginseng Grey Mold Resistant to Carbendazim and the Mixture of Carbendazin Plus Diethofencarb

  • Kim, Joo-Hyung;Min, Ji-Young;Bae, Young-Seok;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.322-327
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    • 2009
  • A total of 23 isolates of Botrytis cinerea causing the grey mold were collected from infected ginseng in several fields of Korea. The sensitivity to carbendazim and the mixture of carbendazim plus diethofencarb was determined through a mycelial inhibition test on PDA amended with or without fungicides. B. cinerea isolates were classified as 3 phenotypes, which were the first phenotype resistant to both of carbendazim and the mixture ($Car^RMix^R$), the second one resistant to carbendazim and sensitive to the mixture ($Car^RMix^S$), and the last one sensitive to both of them ($Car^RMix^S$). Carbendazim resistance correlated with a single mutation $\beta$-tubulin gene of B. cinerea amplified with primer pair tubkjhL and tubkjhR causing a change of glutamate to alanine at amino acid position 198. Furthermore, the substitution of valine for glutamate led the resistance to carbendazim and the mixture at the same position of amino acid. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis using the restriction endonuclease, Tsp451 and BstUI allowed differentiation of the PCR fragment of $\beta$-tubulin gene of $Car^SMix^S$ isolates from that of $Car^RMix^R$ and $Car^RMix^S$ isolates. This method will aid in a fast detection of resistance of carbendazim and the mixture of carbendazim plus diethofencarb in B. cinerea in ginseng field.

Identification and Characterization of Aspergillus oryzae Isolated from Soybean Products in Sunchang County (순창군 장류로부터 분리된 황국균의 동정 및 특성)

  • Lim, Eunmi;Lee, Ji Young;Elgabbar, Mohammed A. Abdo;Han, Kap-Hoon;Lee, Bo-Soon;Cho, Yong Sik;Kim, Hyoun-Young
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.282-288
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    • 2014
  • In this study, we attempted to isolate fungi from soybean fermented foods produced in Sunchang County and to identify Aspergillus oryzae from fungal isolates. Ten fungal isolates were identified with ${\beta}$-tubulin gene. According to the sequences of ${\beta}$-tubulin gene, ten fungal isolates were identified as A. oryzae/flavus complex. For further identification of the ten of fungal isolates, omtA gene, one gene of the aflatoxin biosynthesis gene cluster, was sequenced and the sequences were compared with those of A. oryzae and A. flavus strains from the GenBank database. In addition, identification of the ten fungal isolates was further confirmed using the PCR amplicon of norB and cypA intergenic region, in which a deletion was recognized relative to A. flavus and A. parasiticus. The amplicon size of the ten fungal isolate strains was smaller than those of A. flavus and A. parasiticus, but the same as that of the reference A. oryzae strain. These results indicated that the ten isolates should be identified as A. oryzae. The protease activity in rice koji made with 6, 13, 17, 27, 37 and 38 of strain, respectively was twice higher than that in control. The kojis made with nine of the A. oryzae isolates, respectively, did not produce aflatoxin, suggesting that the strains could possibly be used as starters for soybean products.

The Role of Cytoskeletal Elements in Shaping Bacterial Cells

  • Cho, Hongbaek
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.307-316
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    • 2015
  • Beginning from the recognition of FtsZ as a bacterial tubulin homolog in the early 1990s, many bacterial cytoskeletal elements have been identified, including homologs to the major eukaryotic cytoskeletal elements (tubulin, actin, and intermediate filament) and the elements unique in prokaryotes (ParA/MinD family and bactofilins). The discovery and functional characterization of the bacterial cytoskeleton have revolutionized our understanding of bacterial cells, revealing their elaborate and dynamic subcellular organization. As in eukaryotic systems, the bacterial cytoskeleton participates in cell division, cell morphogenesis, DNA segregation, and other important cellular processes. However, in accordance with the vast difference between bacterial and eukaryotic cells, many bacterial cytoskeletal proteins play distinct roles from their eukaryotic counterparts; for example, control of cell wall synthesis for cell division and morphogenesis. This review is aimed at providing an overview of the bacterial cytoskeleton, and discussing the roles and assembly dynamics of bacterial cytoskeletal proteins in more detail in relation to their most widely conserved functions, DNA segregation and coordination of cell wall synthesis.

Twig Blight on Chinese Magnolia Vain Caused by Botryosphaeria dothidea in Korea (Botryosphaeria dothidea에 의한 오미자 줄기마름병)

  • Park, Sangkyu;Kim, Seung-Han;Lee, Seung-Yeol;Back, Chang-Gi;Kang, In-Kyu;Jung, Hee-Young
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.44-49
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    • 2016
  • The twig blight symptoms were observed in Chinese magnolia vine (Schisandra chinensis) at Mungyeong city, Gyeongbuk province, Korea in June 2015. The typical symptoms of infected plant were shriveled and wilted in leaves which led to blight resulted in death. Based on the morphological characteristics, the isolate was suspected as Botryosphaeria sp. Inoculation of isolated pathogen was performed to identify its pathogenicity according to Koch's postulates. Re-isolated fungi from the inoculated stem was showed same morphological characteristics with original pathogen. Phylogenetic analysis was performed using combined sequence of rDNA internal transcribed spacer region, EF1-${\alpha}$ and ${\beta}$-tubulin gene. The isolated pathogen was identified to the B. dothidea by phylogenetic analysis. This is the first report of twig blight on S. chinensis caused by B. dothidea in Korea.

Neural and Cholinergic Differentiation of Mesenchymal Stem Cells Derived from the Human Umbilical Cord Blood (인간 제대혈액에서 유래된 중간엽 줄기세포의 신경 및 콜린성 분화)

  • Kam, Kyung-Yoon;Kang, Ji-Hye;Do, Byung-Rok;Kim, Hea-Kwon;Kang, Sung-Goo
    • Development and Reproduction
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    • v.11 no.3
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    • pp.235-243
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    • 2007
  • Human umbilical cord blood(HUCB) contains a rich source of hematopoietic stem cells, mesenchymal stem cells and endothelial cell precursors. Mesenchymal stem cells(MSCs) in HUCB are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. We studied on transdifferentiation-promoting conditions in neural cells and cholinergic neuron induction of HUCB-derived MSCs. Neural differentiation was induced by addingdimethyl sulphoxide(DMSO) and butylated hydroxyanisole(BHA) in Dulbeco's Modified Essential Medium(DMEM) and fetal bovine serum(FBS). Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor(bFGF), retinoic acid(RA) and sonic hedgehog(Shh). MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including $\beta$-tubulin III, GFAP and MBP, was markedly elevated during this acute differentiation. The differentiation rate was about $32.3{\pm}2.9%$ for $\beta$-tubulin III-positive cells, $11.0{\pm}0.9%$ for GFAP, and $9.4{\pm}1.0%$ for Gal-C. HUCB-MSCs treated combinatorially with bFGF, RA and Shh were differentiated into cholinergic neurons. After cholinergic neuronal differentiation, the $\beta$-tubulin III-positive cell population of total cells was $31.3{\pm}3.2%$ and of differentiated neuronal population, $70.0{\pm}7.8%$ was ChAT-positive showing 3 folds higher in cholinergic population than neural induction. Conclusively, HUCB-derived MSCs can be differentiated into neural and cholinergic neurons and these findings suggest that HUCB are alternative cell source of treatment for neurodegenerative diseases such as Alzheimer's disease.

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Proteomic Changes by Acupuncture Stimulation at HT7 in the Hippocampus of Rat Pups (신문혈 자침이 어린 백서 해마의 단백질 발현에 미치는 영향)

  • Bae, Chang-Hwan;Kim, Dong-Soo;Kim, Seung-Tae
    • Korean Journal of Acupuncture
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    • v.29 no.2
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    • pp.260-270
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    • 2012
  • Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

Identification of Genes Differentially Expressed in the MCF-7 Cells Treated with Mitogenic Estrogens

  • Cheon, Myeong-Sook;Yoon, Tae-Sook;Lee, Do-Yeon;Choi, Go-Ya;Lee, A-Yeong;Choo, Byung-Kil;Kim, Ho-Kyoung
    • Journal of Applied Biological Chemistry
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    • v.51 no.1
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    • pp.1-6
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    • 2008
  • Estrogens, a group of steroid compounds functioning as the primary female sex hormone, play an important role in the development and progression of breast cancer. In this study, using a novel annealing control primer-based GeneFishing PCR technology, five differentially expressed genes (DEGs), expressed using 10nM mitogenic estrogens, $17{\beta}$-estradiol (E2) and $16{\alpha}$-hydroxyestrone ($16{\alpha}$-OHE1), were selected from the estrogen receptor (ER)-positive MCF-7 human breast cancer cells. The DEGs, MRPL42, TUBA1B, SSBP1, KNCT2, and RUVBL1, were identified by comparison with the known genes via direct sequencing and sequence homology search in BLAST. Quantitative real-time PCR data showed that two DEGs, tubulin ${\alpha}1b$ and kinetochore associated 2, were greater than 2-fold upregulated by E2 or $16{\alpha}$-OHE1. Both genes could be new biomarkers for the treatment and prognosis of cancers, and further study may provide insights into the molecular mechanisms underlying development and progression of breast cancer.

Regulation of Apoptosis and Cell Cycle in Irradiated Mouse Brain (마우스의 대뇌조직에서 방사선에 의한 아포토시스와 세포주기의 조절)

  • Oh, Won-Yong;Song, Mi-Hee;Chung, Eun-Ji;Seong, Jin-Sil;Suh, Chang-Ok
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.146-152
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    • 2001
  • Purpose : To investigate the regulation of apoptosis and cell cycle in mouse brain irradiation. Materials and Methods : 8-week old male mice, C57B1/6J were given whole body $\gamma-radiation$ with a single dose of 25 Gy using Cobalt 60 irradiator. At different times 1, 2, 4, 8 and 24hr after irradiation, mice were killed and brain tissues were collected. Apoptotic cells were scored by TUNEL assay. Expression of p53, Bcl-2, and Bax and cell cycle regulating molecules; cyclins Bl, Dl, E and cdk2, cdk4, $p34^{cdc2}$ were analysed by Western blotting. Cell cycle was analysed by Flow cytometry. Results : The peak of radiation induced apoptosis is shown at 8 hour after radiation. With a single 25 Gy irradiation, the peak of apoptotic index in C57B1/6J is $24.0{\pm}0.25$ (p<0.05) at 8 hour after radiation. Radiation upregulated the expression of p53/tubulin, Bax/tubulin, and Bcl-2/tubulin with 1.3, 1.1 and 1.45 fold increase, respectively were shown at the peak level at 8 hour after radiation. The levels of cell cycle regulating molecules after radiation are not changed significantly except cyclin D1 with 1.3 fold increase. Fractions of Go-Gl, G2-M and S phase in the cell cycle does not specific changes by time. Conclusion : In mouse brain tissue, radiation induced apoptosis is particularly shown in a specific area, subependyma. These results and lack of radiation induced changes in cell cycle ofter better understanding of radiation response of noraml brain tissue.

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Effects of $Taxol^{TM}$ and Cytochalasin B on the Developmental Capacity of Vitrified Porcine Immature Oocytes

  • Kim, S. W;H. T. Cheong;B. K. Yang;Kim, C. I.;Park, C. K.
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.199-199
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    • 2004
  • This study was conducted to investigate cytoskeleton alterations during vitrified (Open Pulled Straw method) porcine immature oocytes, to utilize Taxol/sup TM/ (polymerization of tubulin molecules) and Cytochalasin B (CB, depolymerization of actin filaments) during vitrification to stabilize microtubule and microfilaments (MT and MF), and to determine in vitro maturation, fertilization and development of cytoskeletal-stabilized and vitrified porcine immature oocytes. (omitted)

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Taxonomical Study of Noteworthy Species of Botryosphaeria in Japan

  • Hattori, Yukako;Ando, Yuho;Sasaki, Atsuko;Uechi, Nami;Nakashima, Chiharu
    • Mycobiology
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    • v.49 no.2
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    • pp.122-132
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    • 2021
  • The reexamination of the fungal genus Botryosphaeria on 12 plant species of 10 families was carried out based on molecular phylogenetic analyses using the regions of translation elongation factor 1-α, β-tubulin, DNA-directed RNA polymerase II subunit, and internal transcribed spacer region of rDNA and morphological characteristics. Japanese isolates were divided into five clades and include Botryosphaeria dothidea, B. qingyuanensis, B. sinensis, and Botryosphaeria spp. Two species, B. qingyuanensis and B. sinensis have been newly added to the Japanese mycoflora, but their host plants are not specified. Botryosphaeria tenuispora isolated from Leucothoe fontanesiana and insect galls on fruits of Aucuba japonica has been proposed as a new species.