• Title/Summary/Keyword: $\alpha$-tubulin

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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.

Analysis of Housekeeping Gene Expression in Mice Administered to GM and non-GM Cabbage (유전자변형 배추를 섭취한 마우스 장기에서의 Housekeeping Gene의 발현 분석)

  • Lee, Dong-Youb;Heo, Jin-Chul;Kim, Kyung-Hae;Han, Song-Yi;Cho, Hyun-Seok;Lee, Sang-Han
    • Food Science and Preservation
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    • v.15 no.1
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    • pp.84-87
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    • 2008
  • We used RT-PCR to measure housekeeping gene expression in mice fed GM and non-GM cabbage, in an effort to evaluate the risk of GM food to humans. After normalization of housekeeping gene levels, highly uniform expression may be seen in many organisms during various stages of development and under different environmental conditions. We assessed the expression of four genes in Chinese cabbage; these were Profilin, Tubulin-alpha (Tub-1), Heat-shock protein (Bchsp 17.6), and Ubiquitin conjugating enzyme (UBE). We measured the expression of four well-known housekeeping genes in mice: ${\beta}$-actin, (${\beta}$-act), ${\beta}$-2-microglobulin(B2m), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ${\beta}$-glucuronidase (Gus). Gene expression was measured in liver, stomach, small intestine, large intestine, kidney, and spleen of mice fed GM or non-GM cabbage. No significant expression differences were found.

Phospholipase C-γ Activation by Direct Interaction with β-Tubulin Isotypes (베타 튜불린에 의한 포스포리파제 C-감마1의 활성화)

  • Lee, In-Bum;Kim, Sung-Kuk;Choi, Jang-Hyun;Suh, Pann-Ghill;Chang, Jong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.612-617
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    • 2006
  • Phosphoinositide-specific phospholipase $C-{\gamma}\;1\; (PLC-{\gamma}\;1)$ has pivotal roles in cellular signaling by producing second messengers, inositol 1,4,5-trisphosphate $(IP_3)$ and diacylglycerol (DG). Tubulin is a main component of microtubules and mitotic spindle fibers, which are composed of ${\alpha}-$ and ${\beta}-tubulin$ heterodimers in all eukaryotic cells. In humans, six ${\beta}-tubulin$ isotypes have been identified which display a distinct pattern of tissue expression. Previously we found that $PLC-{\gamma}\;1$ and one of four ${\beta}-tubulin$ isotypes including ${\beta}1$, ${\beta}2$, ${\beta}3$ and ${\beta}6$, colocalized in COS-7 cells and cotranslocated to the plasma membrane to activate $PLC-{\gamma}\;1$ upon agonist stimulation. In the present study, we demonstrate that the remaining two, tubulin ${\beta}4$ and ${\beta}5$, also showed a potential to activate $PLC-{\gamma}\;1$. The phosphatidylinositol 4,5-bisphosphate $(PIP_2)$ hydrolyzing activity of $PLC-{\gamma}\;1$ was substantially increased in the presence of purified ${\beta}4$ and ${\beta}5$ tubulin in vitro, whereas the activity was not promoted by bovine serum albumin, suggesting that tubulin ${\beta}4$ and ${\beta}5$ also activate $PLC-{\gamma}\;1$. Taken together, our results suggest that all the ${\beta}-tubulin$ isotype activates $PLC-{\gamma}\;1$ activity to regulate cellular signaling.

A Tubulin Inhibitor, N-(5-Benzyl-1,3-thiazol-2-yl)-3-(furan-2-yl)prop-2-enamide, Induces Anti-inflammatory Innate Immune Responses to Attenuate LPS-mediated Septic Shock

  • Park, Hyun Jung;Lee, Sung Won;Park, Hwangseo;Park, Se-Ho;Hong, Seokmann
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3307-3312
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    • 2014
  • The anti-inflammatory effect of a tubulin inhibitor, N-(5-benzyl-1,3-thiazol-2-yl)-3-(furan-2-yl)prop-2-enamide (1), on innate immune responses remains unclear. Thus, we investigated the effect of 1 on the immune responses mediated by lipopolysaccharide (LPS). The in vitro addition of 1 to dendritic cells and macrophages dose-dependently reduced tumor necrosis factor alpha production elicited by LPS stimulation. Additionally, the stimulation of natural killer (NK) and natural killer T (NKT) cells with 1 resulted in the decrease of interferon gamma ($IFN{\gamma}$) induced by LPS treatment. Moreover, 1 substantially reduced interleukin 12 in dendritic cells (DC) as well as $IFN{\gamma}$ in NKDCs induced by LPS in vitro. Furthermore, the in vivo administration of 1 ameliorated LPS/D-galactosamine-induced endotoxic lethality in mice. Taken together, our results demonstrate for the first time that 1 possesses anti-inflammatory properties, most notably by modulating LPS-induced innate immune responses. Therefore, 1 might have therapeutic potential for the treatment of inflammation-mediated diseases such as sepsis.

Unrecorded Fungal Species Isolated from Indoor Air in the Log Bed- and Sawdust Media-based Mushroom Cultivation Houses (원목 및 톱밥배지 버섯 재배사 내 실내 공기서 분리한 미기록 진균 보고)

  • Ahn, Geum Ran;Kim, Ji Eun;Kim, Jun Young;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.495-503
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    • 2018
  • Oak mushroom is cultivated using logs and sawdust media as substrates. In this study, fungi were isolated during a monitoring of indoor air in the oak mushroom cultivation houses located in Cheongyang-gun of Chungnam, Geoje-gun of Gyeongnam, Gumi-si of Gyeongbuk, Jangheung-gun of Jeonnam and Yeoju-si of Gyeongggi-do. Identification of the fungi based on morphology and molecular analysis of the internal transcribed spacer region and 28S rDNA, translation elongation factor translation elongation factor 1 a gene, and ${\beta}-tubulin$ gene revealed that six fungi, Cenangium acuum, Neopestalotiopsis surinamensis, Metarhizium marquandii, Periconia macrospinosa, Trichoderma petersenii, and Trichoderma paratroviride that have not been recorded previously in Korea.

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.

First Report of Peach Fruit Rot Caused by Fusarium avenaceum in Korea (Fusarium avenaceum에 의한 복숭아 신규 과실 썩음병 발생 보고)

  • Heo, A Yeong;Koo, Young Mo;Choi, Young-Joon;Kim, Sang Hee;Chung, Gyu Young;Choi, Hyong Woo
    • Research in Plant Disease
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    • v.26 no.1
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    • pp.48-52
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    • 2020
  • In July 2019, typical rot symptom was observed on peach fruits harvested from the fields at Andong, Korea. As the disease progressed, white and purple colored mycelial mat developed on the surface of the infected fruits. A causal pathogen was isolated from the infected fruit and cultured on potato dextrose agar media for identification. Fungal colonies on potato dextrose agar produced 3 pigments, including purple, yellow, and white colors. The isolate incited fruit rot symptoms on artificially inoculated peach fruits, from which the same fungus was isolated, fulfilling Koch's postulates. Based on the morphological characteristics and sequence analysis of rDNA internal transcribed spacer, translation elongation factor 1-alpha, and β-tubulin, the causal agent of the disease was identified as Fusarium avenaceum. This study is the first report of fruit rot of peach fruits caused by Fusarium avenaceum in Korea.

Keeping house: evaluation of housekeeping genes for real-time PCR in the red alga, Bostrychia moritziana (Florideophyceae)

  • Shim, Junbo;Shim, Eunyoung;Kim, Gwang Hoon;Han, Jong Won;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.31 no.2
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    • pp.167-174
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    • 2016
  • Biological response of cells to variable conditions should affect the expression level of certain genes. Quantification of these changes in target genes needs stable internal controls. Real-time quantitative polymerase chain reaction (PCR) has traditionally used reference or ‘housekeeping’ genes, that are considered to maintain equal expression in different conditions, to evaluate changes in target genes between samples and experimental conditions. Recent studies showed that some housekeeping genes may vary considerably in certain biological samples. This has not been evaluated in red algae. In order to identify the optimal internal controls for real-time PCR, we studied the expression of eleven commonly used housekeeping genes; elongation factor 1-alpha, glyceraldehyde-3-phosphate dehydrogenase, β-actin, polyubiquitin, 30S ribosomal gene, 60S ribosomal gene, beta-tubulin, alpha-tubulin, translation initiation factor, ubiquitin-conjugating enzyme, and isocitrate dehydrogenase in different life-history stages of Bostrychia moritziana. Our results suggest that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 30S ribosomal gene, have the most stable gene expression levels between the different life history stages (male, female, carposporophyte, and tetrasporophyte), while the other genes are not satisfactory as internal controls. These results suggest that the combinations of GAPDH and 30S would be useful as internal controls to assess expression level changes in genes that may control different physiological processes in this organism or that may change in different life history stages. These results may also be useful in other red algal systems.