• Environmental Analysis Health and Toxicology
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• v.22 no.4
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• pp.305-312
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• 2007

환경오염을 신속하게 모니터링하기 위한 생물지표의 개발은 증가하고 있는 오염의 심각성에 비추어 매우 중요한 과제로 여겨지고 있다. 본 연구에서는 독성물질처리에 의하여 선택적으로 발현이 조절되는 단백질의 동정을 통하여 독성물질에 대한 단백질 생물지표를 발굴하고자 시도하였다. 즉, 송사리(Oryzias latipes)를 유기인계 살충제인 다이아지논(diazinon)에 0, 0.1, 1, 5 mg/L 농도로 24시간 노출시킨 후, 머리와 몸통부분으로 나누어 단백질 발현패턴을 분석하였다. 본 시스템에서 다이아지논 처리에 의하여 유의적으로 발현이 증가된 단백질로서 alpha-tubulin, ribonuclease pancreatic precursor, protein hfq 등을 동정하였으며, 이 가운데 alpha-tubulin과 $hsp90{\beta}$의 발현이 다이아지논 농도에 의존적으로 증가하는 것을 semi-quantitative RT-PCR방법으로 확인하였다. 이와 같이 다이아지논 처리에 특이적으로 발현이 증가된 송사리 단백질들은 노출평가를 위한 생물지표로서 개발에 응용될 수 있을 것으로 평가된다.

• Toxicological Research
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• v.20 no.1
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• pp.21-29
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• 2004

We used cDNA microarray to assess gene expression profiles in hematopoetic cell line, U-937, exposed to low doses of ionizing irradiation. The 1,000 DNA elements on this array were PCR-amplified cDNAs selected from named human cancer related genes. According to the strength of irradiation, the levels of some gene expression were increased or decreased as dose-dependent manner. The gene expressions of Tubulin alpha, protein kinase, interferon-alpha, -beta, -omega receptor and ras homolog gene family H were significantly increased. Especially, Tubulin gene was shown 2.5 fold up-regulated manner under stress of 500 rad irradiation than 200 rad. On the other hand, fibroblast growth factor 12 and four and a half LIM domains, etc. were significantly down-regu-lated. Also, tumor protein 53(TP53) related genes that p53 inducible protein, tumor protein 53-binding protein looks of little significance as radiation sensitive manner. The radio-sensitivity of tubulin gene etc. that we proposed could be useful to rapid and correct survey for the bio-damage by exposure to low dose irradiation.

• 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.

• 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.

• The Plant Pathology Journal
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• v.24 no.3
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• pp.269-278
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• 2008

Ripe rot was frequently observed on fruits, leaves and stems of grape growing in eight locations in Korea from 2004 to 2006. All 30 isolates of Colletotrichum sp. were obtained from lesions of the ripe rot on grape plants. Out of the isolates, 19 isolates were identified as Colletotrichum acutatum and the others as Colletotrichum gloeosporioides based on morphological and cultural characteristics. Inter and intra specific variations of the Colletotrichum spp. isolates were investigated using RAPD and sequences of rDNA ITS and $\beta$-tubulin-2. Isolates of C. acutatum and C. gloeosporioides were distinctly differentiated by molecular analyses. Phylogenetic trees of ITS and$\beta$-tubulin-2 showed that Korean isolates of C. acutatum were clustered into groups A2 and A3 among the eight global groups. A2 included non-chromogenic isolates and A3 chromogenic ones. Both C. acutatum and C. gloeosporioides isolates were tested for pathogenicity to grape leaves. All isolates tested induced lesions on the leaves of grape by artificial inoculation. There was no difference in pathogenicity between C. acutatum and C. gloeosporioides isolates. This is the first report that C. acutatum except C. gloeosporioides causes grape ripe rot in Korea.

• 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.

• Journal of Korean Neurosurgical Society
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• v.44 no.6
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• pp.375-381
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• 2008

Objective : The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF). insulin growth factor-I (IGF-I). brain-derived neurotrophic factor (BDNF). and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. Methods : Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using $\beta$-tubulin III and NeuN as neural markers. Results : Neural differentiations in the presence of individual growth factors for $\beta$-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-I, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for $\beta$-tubulin III and 3.1% for NeuN. For $\beta$-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1 %; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). Conclusion : Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and post-mitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.

• 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.

• Mycobiology
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• v.35 no.4
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• pp.180-185
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• 2007

Grape fruits with blue mold symptoms were collected from house storages in different locations in Korea and were investigated for their association with Penicillium species. A total of 12 isolates of Penicillium were isolated from the collected fruits. Based on morphological and cultural characteristics and ${\beta}-tublin$ gene sequence data analysis, they were identified as P. bialowiezense, P. citrinum, P. echinulatum, P. expansum, P. solitum and unidentified Penicillium species. P. solitum was the predominant followed by P. expansum. P. bialowiezense and P. echinulatum were newly recorded in Korea. ${\beta}-Tubulin$ gene sequences could be used to distinguish each species of Penicillium and the molecular groups were correlated well with the morphological species. The unidentified species was supposed to be a new species, not previously reported in literature.

• Journal of Life Science
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• v.29 no.2
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• pp.256-264
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• 2019

Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. In contrast, MLCK and ROCK play critical roles in the regulation of stress fiber (SF) formation in cells. To determine whether $LT{\beta}R$ stimulation in fibroblastic reticular cells (FRCs) is involved in these signaling pathways, myosin light chain kinase inhibitor-7 (ML-7) was used to inhibit them. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic anti-$LT{\beta}R$ antibody-treated cells. The inhibition of ROCK activity with Y27632-induced changes in actin cytoskeleton organization and cell morphology in FRCs. Actin bundles rearranged into SFs, and phospho-myosin light chain (p-MLC) co-localized in FRCs. We checked the level of Rho-guanosine diphosphate (RhoGDP)/guanosine triphosphate (GTP) exchange activity using FRC lysate. When $LT{\beta}R$ was stimulated with agonistic anti-$LT{\beta}R$ antibodies, Rho-GDP/GTP exchange activity was markedly reduced. Regarding $LT{\beta}R$ signaling with a focus on MLCK inhibition, we showed that the phosphorylation of MLCs was reduced by $LT{\beta}R$ stimulation in FRCs. Cytoskeleton components, such as tubulin, b-actin, and phospho-ezrin proteins acting as membrane-cytoskeleton linkers, were produced in de-phosphorylation, and they reduced expression in agonistic anti-$LT{\beta}R$ antibody-treated FRCs. Collectively, the results suggested that MLCK and ROCK were simultaneously responsible for SF regulation triggered by $LT{\beta}R$ signaling in FRCs.