• Title/Summary/Keyword: Beta-tubulin

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Fungal Diversity and Enzyme Activity Associated with the Macroalgae, Agarum clathratum

  • Lee, Seobihn;Park, Myung Soo;Lee, Hanbyul;Kim, Jae-Jin;Eimes, John A.;Lim, Young Woon
    • Mycobiology
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    • v.47 no.1
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    • pp.50-58
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    • 2019
  • Agarum clathratum, a brown macroalgae species, has recently become a serious environmental problem on the coasts of Korea. In an effort to solve this problem, fungal diversity associated with decaying A. clathratum was investigated and related ${\beta}$-glucosidase and endoglucanase activities were described. A total of 233 fungal strains were isolated from A. clathratum at 15 sites and identified 89 species based on morphology and a multigene analysis using the internal transcribed spacer region (ITS) and protein-coding genes including actin (act), ${\beta}$-tubulin (benA), calmodulin (CaM), and translation elongation factor (tef1). Acremonium, Corollospora, and Penicillium were the dominant genera, and Acremonium fuci and Corollospora gracilis were the dominant species. Fifty-one species exhibited cellulase activity, with A. fuci, Alfaria terrestris, Hypoxylon perforatum, P. madriti, and Pleosporales sp. Five showing the highest enzyme activities. Further enzyme quantification confirmed that these species had higher cellulase activity than P. crysogenum, a fungal species described in previous studies. This study lays the groundwork for bioremediation using fungi to remove decaying seaweed from populated areas and provides important background for potential industrial applications of environmentally friendly processes.

CD Gene Microarray Profiles of Bambusae Caulis in Liquamen in Human Mast Cell

  • Jeon Hoon;Kang Nan Joo;Kim Gyo Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.1
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    • pp.241-246
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    • 2003
  • Bambusae Caulis in Liquamen(BCL) has been used to relieve the cough and asthma, and remove the phlegm in traditional Oriental medicine. In recent years, it was studied for its antiinflammatory, antiallergenic, immune-modulating, and anticarcinogenic capabilities. This experiment was performed to evaluate the microarray profiles of CD genes in human mast cells before and after BCL treatment. The results are as follows: The expression of 51 of the genes studied was up-regulated in the Bel-treated group; they include the genes coding L apoferritin, beta-2-microglobulin, ferritin light polypeptide, CD63, monocyte chemotactic and activating fact, heme oxygenase 1, CD140a, integrin alpha M, colony stimulating factor 2 receptor, eukaryotic translation elongation factor, CD37, interleukin 18, NADH dehydrogenase 1 beta, CD48, 5-lipoxygenase activating protein, interleukin 4, ribosomal protein L5, GABA(A) receptor-associated protein, beta-tubulin, integrin beta 1, CD162, CD32, lymphotoxin beta, alpha-tublin, integrin alpha L, CD2, CD151, CD331, 90 kDa heat shock protein, CD59, CD3Z, microsomal glutathione S-transferase 2, CD33, CD162R, cyclophilinA, CD84, interleukin 9 receptor, interleukin 11, CD117, CD39-Like 2, and so forth. The expression of 7 of the genes studied was down-regulated in the BCL-treated group; they include the genes coding con, CD238, SCF, CD160, CD231, CD24, and CD130. Consequently, the treatment of BCL on the human mast cells increased the expression of 51 genes and decreased the expression of 7 genes. These data would provide a fundamental basis to the traditional applications of Bambusae Caulis in Liquamen.

Pepper Blight Disease Inhibition Metagenome Clone Screening Using Soil Metagenome Library (토양 Metagenome Library로부터 고추역병 저해 클론 탐색)

  • Park, Hae-Chul;Sung, So-Ra;Kim, Dong-Gwan;Koo, Bon-Sung;Jeong, Byeong-Moon;Kim, Jin-Heung;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.228-231
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    • 2009
  • We have purified Phytophthora capsici alpha and beta tubulin from Escherchia coli BL21(DE3). The recombinant alpha and beta tubulins were assembled into microtubule in vitro with specific conditions. The metagenome library was isolated from soil in the Mt. Yeo-Ki, Suwon, Korea and manufactured with the method mentioned in experiment contents for in vitro screening of microtubule assembly screening. FRET effect was used for microtubule assembly inhibitor screening with metagenome library. We got 2 metagenome clones from in vitro screening, and these 2 hit clones showed P. capsici growth inhibition activity on the growing pepper plants. These results suggest that new development of potent inhibitor for pepper blight disease and new approach to prevention of pepper blight disease.

Cleavage Pattern of Urechis unicinctus Eggs in Vitro Fertilization (인공수정에 의한 Urechis unicinctus 난자의 난할형식)

  • Shin, Kil-Sang;Lee, Dae-Hee;Ko, Tae-Young
    • Applied Microscopy
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    • v.34 no.1
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    • pp.71-81
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    • 2004
  • When U. unicinctus mature oocytes were fertilized in vitro, germinal vesicle breakdown (GVBD) and meioses occurred and the zygotes entered cleavage stage. A modified pattern of spiral cleavages, suggestively based on behavior of mitotic spindles, have been observed in this work. The first and second cleavages were meridional and the third was equatorial, and then followed by repetitions of meridional-equatorial cleavage. The cleavage of the isolecithal egg were equal and holoblastic and its patterns were spiral. The anti-${\alpha}-,-{\beta}$- tubulin reactions and confocal microscopy revealed mitotic apparates tilted obliquely at each mitosis causing oblique displacements of the blastomeres. Despite isolecithal distribution of yolk, this observations implicated that tilting of mitotic apparates induced spiral cleavage and the displacements of blastomeres. However, these features would not be the typical spiral cleavage, but represented a modified pattern of known Spiralian s in the sense of the equal cleavage. During the first cleavage, heart-shaped eggs have been produced. Electron microscopies exhibited the first cleavage furrow extended with its membranous structure deeply into the cytoplasm. Contractile ring has not been observed.

Comparison of Expression Pattern of Housekeeping Genes in Mice fed Genetically Modified Rice (유전자 이입에 따른 GM쌀 섭취 마우스의 Housekeeping Gene 발현 패턴 비교)

  • Lee, Dong-Yeob;Heo, Jin-Chul;Lee, Kyu-Hyun;Kim, Dong-Ho;U, Sang-Uk;Cho, Hyun-Suk;Lee, Sang-Han
    • Food Science and Preservation
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    • v.14 no.6
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    • pp.688-694
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    • 2007
  • To evaluate the human risk of long-term intake of genetically modified (GM) rice, we carried out RT-PCR of housekeeping genes. Housekeeping genes, which show highly uniform expression in living organisms during various stages of development and under different environmental conditions, were normalized by RT-PCR. We assessed the expression of 10 common housekeeping genes (18s rRNA, 25S rRNA, UBC, UBQ5, UBQ10, ACT11, GAPDH, eEF-$1{\alpha}$, ${\beta}$-TUB, GAPDH, ${\beta}$-actin, B2m, G6pd2, Gyk, Gus, Hprt, Cyclophlin A, Tfrc, ${\alpha}$-tubulin and RPL13A) in the liver, stomach, small intestine, large intestine, kidney and spleen of mice fed GM or non-GM rice. We found no significant differences in the expression of housekeeping genes between the two groups of mice.

First Report on Four Novel Endophytic Fungal Species Isolated from Leaves of Juniperus chinensis var. sargentii in Korea (눈향나무의 잎에서 분리된 4종의 국내 미기록 내생균)

  • Park, Hyeok;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.114-121
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    • 2018
  • We isolated endophytic fungi from the leaves of the conifer Juniperus chinensis var. sargentii inhabiting Mt. Hallasan. The fungal isolates were identified based on phylogenetic analysis using nucleotide sequences of the internal transcribed spacer region, large subunit region of ribosomal DNA, translation elongation factor, and ${\beta}$-tubulin region. In this study, we confirmed four species of unreported endophytic fungi in Korea, Allantophomopsis lycopodina, Epicoleosporium ramularioides, Kabatina juniperi, and Seiridium podocarpi. We describe their morphological characteristics and the results of the phylogenetic analysis.

Revision of the Lichen Genus Stereocaulon (Stereocaulaceae, Ascomycota) in South Korea

  • Park, Jung Shin;Park, Chan-Ho;Park, Sook-Young;Oh, Soon-Ok;Jayalal, Udeni;Hur, Jae-Seoun
    • Mycobiology
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    • v.46 no.2
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    • pp.101-113
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    • 2018
  • Lichen genus Stereocaulon (Schreb.) Hoffm is distributed throughout the world. Although 15 Stereocaulon species have been recorded in Korea, no detailed taxonomic or revisionary research has been conducted for nearly two decades. In this study, we collected 260 putative Stereocaulon spp. samples and identified the species based on morphological, chemical, and molecular characteristics. From the collected samples, 10 species of Stereocaulon were identified, nine of which had already been reported, although this was the first report for the tenth, S. octomerellum Hue, in Korea. General characteristics of Stereocaulon spp. include coralloid phyllocladia and tubercular cephalodia; however, the specimen first collected in Korea was a rare species with tomentum on the pseudopodetia. The specimen of S. octomerellum is characterized by the presence of a primary thallus, granule to short coralloid phyllocladia, and pseudopodetia up to 1 cm in size, with tubercular cephalodia. To determine the phylogeny of the specimens, we compared the ITS sequences of ribosomal DNA and the b-tubulin gene sequences. Phylogenetic analysis showed that the Korean Stereocaulon species were monophyletic and placed in the previous phylogenetic classification. Species of S. intermedium and S. exutum, however, were polyphyletic, and are morphologically variable and widespread species. Overall, we present here detailed morphological and chemical descriptions of each species identified and a revised key of all known Stereocaulon species in South Korea.

Neuron-specific expression of p48 Ebp1 during murine brain development and its contribution to CNS axon regeneration

  • Ko, Hyo Rim;Hwang, Inwoo;Ahn, So Yoon;Chang, Yun Sil;Park, Won Soon;Ahn, Jee-Yin
    • BMB Reports
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    • v.50 no.3
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    • pp.126-131
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    • 2017
  • P48 Ebp1 is expressed in rapidly proliferating cells such as cancer cells and accelerates cell growth and survival. However, its expression pattern and role in central nervous system development have not been studied. Here, we demonstrated the spatiotemporal expression pattern of p48 Ebp1 during embryonic development and the postnatal period. During embryonic development, p48 Ebp1 was highly expressed in the brain. Expression gradually decreased after birth but was still more abundant than p42 expression after birth. Strikingly, we found that p48 Ebp1 was expressed in a cell type specific manner in neurons but not astrocytes. Moreover, p48 Ebp1 physically interacted with beta tubulin but not alpha tubulin. This fits with its accumulation in distal microtubule growth cone regions. Furthermore, in injured hippocampal slices, p48 Ebp1 introduction promoted axon regeneration. Thus, we speculate that p48 Ebp1 might contribute to microtubule dynamics acting as an MAP and promotes CNS axon regeneration.

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.

Detection of Colletotrichum spp. Resistant to Benomyl by Using Molecular Techniques

  • Dalha Abdulkadir, Isa;Heung Tae, Kim
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.629-636
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    • 2022
  • Colletotrichum species is known as the major causal pathogen of red pepper anthracnose in Korea and various groups of fungicides are registered for the management of the disease. However, the consistent use of fungicides has resulted in the development of resistance in many red pepper-growing areas of Korea. Effective management of the occurrence of fungicide resistance depends on constant monitoring and early detection. Thus, in this study, various methods such as agar dilution method (ADM), gene sequencing, allele-specific polymerase chain reaction (PCR), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied for the detection of benzimidazole resistance among 24 isolates of Colletotrichum acutatum s. lat. and Colletotrichum gloeosporioides s. lat. The result of the ADM showed that C. gloeosporioides s. lat. was classified into sensitive and resistant isolates to benomyl while C. acutatum s. lat. was insensitive at ≥1 ㎍/ml of benomyl. The sequence analysis of the β-tubulin gene showed the presence of a single nucleotide mutation at the 198th amino acid position of five isolates (16CACY14, 16CAYY19, 15HN5, 15KJ1, and 16CAYY7) of C. gloeosporioides s. lat. Allele-specific PCR and PCR-RFLP were used to detect point mutation at 198th amino acid position and this was done within a day unlike ADM which usually takes more than one week and thus saving time and resources that are essential in the fungicide resistance management in the field. Therefore, the molecular techniques established in this study can warrant early detection of benzimidazole fungicide resistance for the adoption of management strategies that can prevent yield losses among farmers.