• Title/Summary/Keyword: Acidic proteins

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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.

Functional Roles of the Aromatic Residues in the Stabilization of the [$Fe_4S_4$] Cluster in the Iro Protein from Acidithiobacillus ferrooxidans

  • Zeng, Jia;Liu, Qing;Zhang, Xiaojian;Mo, Hongyu;Wang, Yiping;Chen, Qian;Liu, Yuandong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.294-300
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    • 2010
  • The Iro protein is a member of the HiPIP family with the [$Fe_4S_4$] cluster for electron transfer. Many reports proposed that the conserved aromatic residues might be responsible for the stability of the iron-sulfur cluster in HiPIP. In this study, Tyr10 was found to be a critical residue for the stability of the [$Fe_4S_4$] cluster, according to site-directed mutagenesis results. Tyr10, Phe26, and Phe48 were essential for the stability of the [$Fe_4S_4$] cluster under acidic condition. Trp44 was not involved in the stability of the [$Fe_4S_4$] cluster. Molecular structure modeling for the mutant Tyr10 proteins revealed that the aromatic group of Tyr10 may form a hydrophobic barrier to protect the [$Fe_4S_4$] cluster from solvent.

Effects of Taraxaci Herba on Cell Death in Breast Cancer Cells (포공영(蒲公英)이 유방암 세포의 사멸에 미치는 영향)

  • Seo, Kook-Jang;Park, Kyung-Mi;Joh, Sung-Hee;Yang, Seung-Jeong
    • The Journal of Korean Obstetrics and Gynecology
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    • v.28 no.2
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    • pp.1-14
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    • 2015
  • Objectives : This study was designed to investigate the effects of Taraxaci Herba (TH) on cell death in breast cancer cells. Methods : In this experiment, the effects of TH on proliferation rates, cell morphology and growth pattern, intracellular reactive oxygen species (ROS) production. In addition, the effects on nuclear condensation, fragmentation and formation of acidic vesicular organelles (AVO) in MCF-7 cells were also investigated. Finally, autophagy related with protein was observed by using western blot method. Results : TH inhibited proliferation of MCF-7 cells, TH elevated intracellular ROS levels significantly. Treatment with TH did not affect nuclear morphologies such as condensation or fragmentation. On the other hand, TH treatment effectively induced AVO. Finally, one of autophagy related with protein, Microtubule-associated proteins 1A/1B light chain 3A (MAP1LC3A, LC3) level was elevated by treatment with TH. Conclusions : These data indicate that TH is able to be used for patient with breast cancer and mechanisms are involved in autophagy through ROS generation.

The Effects of KongJin-Dan(KJD) on the Alzheimer's Disease Model Induced by CT105 (공진단(拱辰丹)이 CT105로 유도된 Alzheimer's disease 병태(病態)모델에 미치는 영향)

  • Chung, Dae-Kyoo;Hwang, Seon-Mi
    • Journal of Oriental Neuropsychiatry
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    • v.15 no.2
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    • pp.103-118
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    • 2004
  • Objective : This experiment was designed to investigate the effect of KongJin-dan(KJD) on the Alzheimer's disease. Method : The effects of KJD on $LI-1{\beta}$, IL-6, $TNF-{\alpha}$, amyloid precursor proteins(APP), acetylcholinesterase(AChE), glial fibrillary acidic protein(GFAP) mRNA of PC-12 and THP-1 cell treated by CT105 and AChE activity, APP production of PC-12 cell lysate treated by CT105 were investigated, respectively. Results : 1. KJD suppressed $LI-1{\beta}$, IL-6, $TNF-{\alpha}$, APP, AChE, GFAP mRNA in THP-1 and PC-12 cell treated by CT105. 2. KJD suppressed AChE activity and production of APP significantly in cell lysate of PC-12 cell treated by CT105. Conclusions : This study shows that KJD might be usefully applied for prevention and treatment of Alzheimer's disease.

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Enhancement of Erythropoietin Production from Chinese Hamster Ovary(CHO) Cells by Introduction of the Urea Cycle Enzymes, Carbamoyl Phosphate Synthetase I and Ornithine Transcarbamylase

  • Kim, Na-Young;Lee, Yun-Jeong;Kim, Hyung-Jin;Choi, Jung-Ho;Kim, Jung-Kwon;Chang, Kern-Hee;Kim, Jung-Hoe;Kim, Hong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.844-851
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    • 2004
  • Efficient mammalian erythropoietin (EPO)-expression systems are required for therapeutic applications. The accumulation of ammonia is a major problem in the production of recombinant proteins in cultured animal cells. To counter this problem we introduced the first two genes of the urea cycle, carbamoyl phosphate synthetase (CPSI) and ornithine transcarbamylase (OTC), into IBE Chinese Hamster Ovary (CHO) cells by stable transfection. The resulting cell line, CO5, had a higher growth rate and accumulated less ammonia per cell than the parental cell line, IBE. In addition, it produced 2 times more EPO than the parent, and the purified EPO contained a higher proportion of acidic isoforms with approximately 15% more sialic acid.

Immunostimulating Components from the Root of Angelica gigas Nakai (참당귀(Angelica gigas Nakai) 뿌리의 면역증강활성 성분)

  • Ahn, Kyung-Seop;Sim, Woong-Seop;Kim, Hwan-Mook;Han, Sang-Bae;Kim, Ik-Hwan
    • Korean Journal of Pharmacognosy
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    • v.27 no.3
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    • pp.254-261
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    • 1996
  • A potent immuno-stimulating activity was detected from the watersoluble and ethanol-precipitated crude extract (AG-1) of the root of Angelica gigas Nakai. The crude extract was fractionated into two fractions, an acidic AG-2 and a neutral AG-3 fraction by DEAE-cellulose adsorption. The two fractions contained polysaccharides of which M.W. were 10 Kdal and >2,000 Kdal respectively, proteins, and various inorganic components. The immunostimulating activities of two fractions were not reduced by proteinase K, acid or alkali treatment. The polysaccharides obtained from the root of A. gigas were mainly composed of arabinose, galactose, and galacturonic acid. These results indicated that immuno- stimulating components of the root of A. gigas was a kind of pectic polysaccharides or arabinogalactans.

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Novel TGACG-Motif Binding Protein of Soybean

  • Hong, Jong-Chan
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.40-47
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    • 1996
  • The promoters of a variety of plant genes are characterized by the presence of TGACG motif-containing sequences. These genes often exhibit quite diverse expression characteristics and in many case the TGACG-motif has been demonstrated to be essential for expression. Here we report the isolation and characterization of a soybean cDNA that encodes a novel basic/leucine zipper (bZIP) protein, STF1, that specifically interacts with Hex (TGACGTGG) and CRE (TGACGTCA) sequences. This protein contains a bZIP motif at C-teminus and an acidic domain at N-terminus. DNA binding specificities, heterodimer formation, and expression characteristics of STF1 were compared with a soybean TGA1 protein, STGA1. The soybean STF1 interacts with TGACG-sequences containing an ACGT core, while STGA1 requires TGACG as a sufficient binding sequence. The flanking sequences to the TGACG motif affected DNA binding of STF1 siginificantly. The STF1 mRNA is found mainly in dark grown soybean seedling with higher expression in apical and elongating hypocotyl, while STGA1 mRNA is highly abundant in roots of light grown plants. Furthermore, we demonstrate that STF1 heterodimerzes with G-box binding factorss (GBFs) which was not observed with TGA1. The fact that STF1 possesses both distinct DNA binding speficities and heterodimerization properties suggest that STF1 belongs to a new family of plant bZIP proteins which recognize the Hex/CRE motif.

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GFAP IMMUNOREACTIVITY IN SATELLITE CEllS OF TRIGEMINAL GANGLION FOllOWING AXOTOMY OF INFERIOR ALVEOLAR NERVE IN RAT (흰쥐에서 하치조신경 절단에 따른 삼차신경절 위성페포에서 GFAP-IR의 변화)

  • Lee, Chang-Seop;Lee, Sang-Ho;Kim, Heung-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.25 no.1
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    • pp.249-256
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    • 1998
  • Glial fibrillary acidic proteins (GFAP) are a group of intermediate filaments that are distributed in the cytoplasm of glial cells. GFAP immunoreactivity (GFAP-IR) increase after central and peripheral nerve injuries. The purpose of this study was to determine change of GFAP-IR in rat trigeminal ganglion satellite cells following the axotomy of inferior alveolar nerve(IAN). The immunohistochemistry was carried out using the avidin-biotin-peroxidase complex(ABC) method. 1. Control group : Astrocytes in central root of trigeminal ganglion had strong GFAP-IR, but satellite cells of trigeminal ganglion occasionally had GFAP-IR. The patterns of reactivity in satellite cells of trigeminal ganglion were not concenturated in any specific region of trigeminal ganglion. 2. Three day group after IAN axotomy : There were highly GFAP-IR in satellite cells of trigeminal ganglion in mandibular region. GFAP-IR in maxillary and ophthalmic regions were less intense compared to mandibular region. 3. Seven day group after IAN axotomy : GFAP-IR that were increased compared to control group were seen in the mandibular region. But GFAP-IR were less intense compared to three day group. These results suggest that GFAP-IR increase in specific region of trigeminal ganglion following peripheral axotomy. therefore we suppose that GFAP study offer research tool in trigeminal neuralgia.

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Animal lectins: potential receptors for ginseng polysaccharides

  • Loh, So Hee;Park, Jin-Yeon;Cho, Eun Hee;Nah, Seung-Yeol;Kang, Young-Sun
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.1-9
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    • 2017
  • Panax ginseng Meyer, belonging to the genus Panax of the family Araliaceae, is known for its human immune system-related effects, such as immune-boosting effects. Ginseng polysaccharides (GPs) are the responsible ingredient of ginseng in immunomodulation, and are classified as acidic and neutral GPs. Although GPs participate in various immune reactions including the stimulation of immune cells and production of cytokines, the precise function of GPs together with its potential receptor(s) and their signal transduction pathways have remained largely unknown. Animal lectins are carbohydrate-binding proteins that are highly specific for sugar moieties. Among many different biological functions in vivo, animal lectins especially play important roles in the immune system by recognizing carbohydrates that are found exclusively on pathogens or that are inaccessible on host cells. This review summarizes the immunological activities of GPs and the diverse roles of animal lectins in the immune system, suggesting the possibility of animal lectins as the potential receptor candidates of GPs and giving insights into the development of GPs as therapeutic biomaterials for many immunological diseases.

Effects of Electron-Beam Irradiation on Saponins and Physio-chemical Properties of Ginseng Powders (Electron Beam 조사가 인삼분말의 사포닌 및 생리화학적 특성에 미치는 영향)

  • 이미경;이종원;도재호;권중호
    • Food Science and Preservation
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    • v.10 no.2
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    • pp.206-212
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    • 2003
  • Comparative effects of electron beam and gamma-ray irradiation (25 ∼15 kGy) were investigated on saponin stability and some physiological and chemical properties of white and red ginseng powders. Saponin components were found stable upon irradiation of both energies when determined by TLC and HPLC, after 4 months of storage at room temperature as well as immediately after treatment The contents of total phenolics and acidic polysaccharides of the samples were higher in red ginseng than in white. Polysaccharide contents increased with irradiation doses. Amylase activity of white ginseng was stimulated by irradiation but decreased with the lapse of storage time. There were no apparent differences in electrophoresis patterns of extracted proteins depending on irradiation doses and energy sources.