• Title/Summary/Keyword: Inducible proteins

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Inhibitory Effect of Sargauum fulvellum Ethanolic Extract on LPS-Induced Inflammatory Reaction in RAW 264.7 Mouse Macrophages

  • Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Jeong, Da-Hyun;Ahn, Dong-Hyun
    • Journal of Applied Biological Chemistry
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    • v.56 no.4
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    • pp.249-255
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    • 2013
  • Recently, algae has been considered as a potential anti-inflammatory source due to its distinctive habitat environment exposing to light and high oxygen concentration. In present study, anti-inflammatory effect of brown alga, Sargassum fullvellum ethanol extract (SFEE), was examined. SFEE inhibited not only the production of nitric oxide and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) but also the expression of inducible nitric oxide synthase and cyclooxygenase 2 in LPS-induced RAW 264.7 cells without affecting cell viability. SFEE also suppressed the expression of nuclear factor kappa B (NF-${\kappa}B$), suggesting that SFEE could affect the expression of inflammation related cytokines and proteins through the regulation of NF-${\kappa}B$. Furthermore, formation of edema of the ear was 40% lower in mice treated with the highest dose (250 mg/kg) of SFEE than in the control mice. Thus, our study showed that SFEE may be a potential therapeutic anti-inflammatory drug.

In Vitro Assay of Mammary Gland Tissue Specific hEPO Gene Expression (hEPO 유전자의 유선조직 특이적 발현에 대한 In Vitro 검정)

  • Koo, Bon Chul;Kwon, Mo Sun;Kim, Teoan
    • Reproductive and Developmental Biology
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    • v.40 no.1
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    • pp.7-13
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    • 2016
  • Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

Molecular Characterization of a PR4 Gene in Chinese Cabbage

  • Chung, Sam-Young;Lee, Kyung-Ah;Oh, Kyung-Jin;Cho, Tae-Ju
    • Animal cells and systems
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    • v.9 no.4
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    • pp.239-244
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    • 2005
  • A cDNA clone for a wound- and pathogen-induced gene in Chinese cabbage (Brassica rapa subsp. pekinensis) was isolated and characterized. The cabbage gene, designated BrPR4, encodes a pathogenesis-related protein 4 (PR4) of 140 amino acids. The BrPR4 protein shows high similarity with wound-inducible antifungal proteins of tobacco, potato, barley, and wheat. The BrPR4 gene is locally induced by a nonhost pathogen, Pseudomonas syringae pv. tomato, that elicits a hypersensitive response in Chinese cabbage. Treatment of the cabbage leaves with benzothiadiazole (BTH), methyl jasmonate or ethephon showed that the BrPR4 gene expression is strongly induced by ethylene, but not by methyl jasmonate or BTH. The BrPR4 gene is also activated by wounding. Interestingly, however, the wound-inducible BrPR4 gene expression is repressed by salicylic acid or BTH, suggesting that there is cross-talk between salicylate-dependent and -independent signaling pathways.

A Phosphate Starvation-Inducible Ribonuclease of Bacillus licheniformis

  • Nguyen, Thanh Trung;Nguyen, Minh Hung;Nguyen, Huy Thuan;Nguyen, Hoang Anh;Le, Thi Hoi;Schweder, Thomas;Jurgen, Britta
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1464-1472
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    • 2016
  • The BLi03719 protein of Bacillus licheniformis DSM13 belongs to the most abundant extracellular proteins under phosphate starvation conditions. In this study, the function of this phosphate starvation inducible protein was determined. An amino-acid sequence analysis of the BLi03719-encoding gene showed a high similarity with genes encoding the barnase of Bacillus amyloliquefaciens FZB42 and binase-like RNase of Bacillus pumilus SARF-032. The comparison of the control strain and a BLi03719-deficient strain revealed a strongly reduced extracellular ribonuclease activity of the mutant. Furthermore, this knockout mutant exhibited delayed growth with yeast RNA as an alternative phosphate and carbon source. These results suggest that BLi03719 is an extracellular ribonuclease expressed in B. licheniformis under phosphate starvation conditions. Finally, a BLi03719 mutant showed an advantageous effect on the overexpression of the heterologous amyE gene under phosphate-limited growth conditions.

Differential Display Detection of Acid-inducible Genes from Porphyra yezoensis Thalli (해조류 방사무늬김 (Porphra yezoensis) 엽체로부터 산 유도 유전자의 분리)

  • JIN Long-Guo;KANG Se-Eun;CHOI Jae-Suk;PARK Sun-Mee;PARK Jung-Youn;JIN Duck-Hee;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.4
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    • pp.269-274
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    • 2004
  • Genetic responses of the edible seaweed Porphyra yezoensis tissue to acid shock have been compared using differential display technique. The tissue was challenged in seawater containing $0.05{\%}$ hydrogen chloride (pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs. Total RNA extracted by the LiCl-guanidium method was reverse transcribed and amplified by PCR with arbitrary primers. The amplified fragment responded by the acid shock was selectively isolated from agarose gel and sequenced with DNA auto sequencer. Sequence (1056 bp) of the cDNA contained at least two genes for ASP7K (MW 7418) and ASP5K (MW 5512) proteins.

Development of Industrial Transgenic Plants Using Antioxidant Enzyme Genes (항산화효소 유전자를 이용한 산업용 형질전환식물체 개발)

  • LEE Haeng-Soon;KIM Kee-Yeun;KWON Suk-Yoon;KWAK Sang-Soo
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04a
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    • pp.49-58
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    • 2002
  • Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21s1 century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (Ipomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.

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Analysis of the Expression Patterns of Thymosin β4, Vascular Endothelial Growth Factor, and Hypoxia-Inducible Factor-1α in Various Tumors Using Tissue Microarray (Tissue microarray를 이용한 여러 암에서의 thymosin β4, vascular endothelial growth factor, 및 hypoxia-inducible factor-1α 발현양상 연구)

  • Lee, Bo-Young;Lee, Seung-Hyun;Ahn, Byung-Kwon;Ock, Mee-Sun;Cha, Hee-Jae
    • Journal of Life Science
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    • v.21 no.3
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    • pp.417-423
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    • 2011
  • Thymosin ${\beta}4$ (TB-4) has been reported to play a key role in tumor growth, metastasis and angiogenesis. In addition, TB-4 induced the expression of vascular endothelial growth factor (VEGF) and stabilized the hypoxia-inducible factor (HIF)-$1{\alpha}$ in melanoma cells. Although the importance of thymosin ${\beta}4$ in angiogenesis and metastasis has been proven, there are few studies that show the expression patterns of TB-4, VEGF and HIF-$1{\alpha}$. This study was conducted to analyze the relationship among these proteins in various tumors. Using tissue microarray analysis, we investigated the expression patterns of TB-4, VEGF and HIF-$1{\alpha}$ in various tumors to identify the expression patterns and relationships of these proteins in certain tumors. TB-4 was highly expressed in osteosarcoma, colon adenocarcinoma, esophageal squamous cell carcinoma, kidney and urinary bladder transitional carcinoma, lung cancer, and liver cancer. HIF-$1{\alpha}$ was highly expressed in nasal cavity inverted papilloma, lung cancer, and esophageal squamous cell carcinoma. The expression patterns of TB-4 and HIF-$1{\alpha}$ were almost similar and co-localized. VEGF expression was high in the blood vessels in tumors, but usually not high in the tumors themselves. VEGF was moderately expressed in stomach cancer, liver angiosarcoma, gall bladder adenocarcinoma, and uterus endometrial adenocarcinoma. The expression patterns of VEGF shows similarities in certain tumors including stomach cancer, osteosarcoma, liposarcoma, lung cancer, liver cancer, gall bladder adenocarcinoma, esophageal squamous cell carcinoma, stomach cancer, colorectal carcinoma and renal cell carcinoma. These results suggest that the expression patterns of TB-4, HIF-$1{\alpha}$ and VEGF were co-localized and related to tumorigenesis and angiogenesis of certain tumors.

Effects of Traditional Wine by using Mycelium of Phellinus linteus on the Expression of Inflammation-Related Proteins in Rat Liver (상황버섯 균사체를 이용한 전통주의 장기투여가 흰쥐 간장에 미치는 영향)

  • Choi, Yung-Hyun;Lee, Yong-Tae;Chung, Kyung-Tae;Jeong, Young-Kee;Choi, Byung-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.642-646
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    • 2007
  • It was examined that the effect of fermented traditional wine made by using mycelium of Phellinus linteus (TWPL) on the expression of inflammation-related proteins in rat liver. Levels of aspartate aminotransferase (AST) was significantly increased in the serum of ethanol-treated rats compared to normal. However, the level of AST showed no significant changes in the TWPL-treated rat compared normal. Slight histopathological changes of liver such as cloudy swelling, inflammatory cells infiltration, Kupffer cell reaction were demonstrated in the rats challenged with ethanol compared with normal. Fewer scores of these changes were observed in TWPL-treated rat with recovered glycogen in hepatocytes of whole hepatic lobule. The RT-PCR and Western analysis showed that the expression of inflammatory proteins such as cyclooxygenase-2, inducible nitric oxide synthase, tumor necrosis factor (TNF)-${\alpha}$ were decreased in the TWPL-treated rat compared with ethanol-treated ones. Immunohistochemical analysis showed that the expression of interleukin-lf and TNF-${\alpha}$ tended to decrease in TWPL-treated rat compared with ethanol-treated ones. These results suggest that TWPL may contains some protective agent for alcohol-induced liver injury through a regulating inflammation-related proteins.

Chloroquine Exerts Anti-metastatic Activities Under Hypoxic Conditions in Cholangiocarcinoma Cells

  • Thongchot, Suyanee;Loilome, Watcharin;Yongvanit, Puangrat;Dokduang, Hasaya;Thanan, Raynoo;Techasen, Anchalee;Namwat, Nisana
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.2031-2035
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    • 2015
  • Intra-tumoral hypoxia is an environment that promotes tumor cell migration, angiogenesis and epithelial-mesenchymal transition that accounts for a major mechanism of metastasis. Chloroquine potentially offers a new therapeutic approach with an 'old' drug for effective and safe cancer therapies, as it exerts anti-metastatic activity. We investigated the inhibitory effect of chloroquine on cholangiocarcinoma (CCA) cell migration under cobalt chloride ($CoCl_2$)-stimulated hypoxia. We showed that chloroquine suppressed CCA cell migration under hypoxic-mimicking conditions on exposure to $100{\mu}M$ $CoCl_2$. Moreover, chloroquine stabilized the protein level of prolyl hydroxylase domain proteins (PHD-2) but reduced the levels of hypoxic responsive proteins such as hypoxia-inducible factor (HIF-$1{\alpha}$) and vascular endothelial growth factor (VEGF). It also suppressed epithelial mesenchymal transition (EMT) by increasing the ratio of E-cadherin to N-cadherin under hypoxic conditions. In conclusion, chloroquine can inhibit hypoxia-stimulated metastasis via HIF-$1{\alpha}$/VEGF/EMT which may serve as a useful additional strategy for CCA therapy.

Identification of a host range determinant from Ralstonia solancearum race 3

  • Yeonhwa Jeong;Lee, Seungdon;Ingyu Hwang
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.71.2-71
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    • 2003
  • Ralstonia solancearum infects many solanaceous plants, however race 3 infects only potato and tomato weakly. To identify genes responsible for race specificity of R. solanacearum, we mobilized genomic library of LSD2029 (race 3) into LSD341 (race 1) and inoculated 1,000 transconjugants into hot pepper. One transconjugant that did not induce wilt symptom in hot pepper was isolated. We found that a cosmid clone, pRSl, conferred avirulence to LSD341. By deletion and mutational analyses of pRSl, we found the 0.9-kb PstI/Hindlll fragment carries avirulence functions. We sequenced the fragment and identified one possible open reading frame, a rsal gene, possibly encoding 110 amino acids. The rsal was preceded with a plant-inducible promoter (PIP) box, indicating that the gene might be regulated by HrpB. Interestingly, the promoter region of the rsal homolog in the strain GM11000 (race 1) did not have the PIP box. Rsal did not show any significant homologies with proteins in the database, indicating th e protein is different from the previously reported avirulence proteins. When we mutated the rsal gene by marker-exchange in LSD2029, the mutant was less virulent in potato.

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