• Title/Summary/Keyword: 조직특이 단백질

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Production of Polyclonal Antibodies Specific to Porcine Adipocyte Plasma Membrane Proteins in Sheep (면양을 이용한 돼지 지방제포 원형질막 단백질 특이 항체의 생산)

  • 최창본;이명진;권은진
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.57-63
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    • 1998
  • The objectives of this study were to produce polyclonal antibody to adipocyte plasma membrane (APM) proteins isolated from pig, and to investigate its tissue specificity. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver and spleen were isolated using a self-forming Percoll gradient. Sheep (40kg) was immunized three times at three week interval with the purified APM proteins. Blood was taken from non-immunized sheep (NS) and from immunized sheep at 10 (AS-1), 12 (AS-2), and 14 (AS-3) days after the third immunization. Antisera titers and cross-reactivity against other tissues were determined by enzyme-linked immunosorbent assay (ELISA). Antisera reacted strongly to APM proteins showing detectable amounts of antibody at 1:81,000 dilution. And antisera showed much stronger reactivity to APM proteins than any other tissue plasma membrane proteins. Furthermore, tissue specificity of antisera against APM was reconfirmed by immunoblotting using anti-sheep immunoglobulin G-horseradish peroxidase conjugate as a secondary antibody Antisera to APM proteins showed adipocyte specificity compared with other tissues. In conclusion, polyclonal antibody against APM proteins isolated from pig was developed successfully in our laboratory, and these antisera showed tissue specificity with APM.

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Chorion Gene Expression in the Cellular Differentiation and Accumulation of Chorion Protein of Silkmoth, Bombyx mandarina I. Specific Structures of Egg-shell and Chorion Protein (한국산 멧누에 (Bombyx mandarina)에 있어서 난각유전자의 형질발현. I. 난각구조의 특이성과 Chorion 단백질)

  • 노시갑
    • Korean journal of applied entomology
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    • v.29 no.3
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    • pp.157-164
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    • 1990
  • The surface patterns and the structures of transverse section of the egg-shell of the sikmoth, Bombyx mandarina, have been described by scanning electron microscope. Three spatially differentiated cross section, called lamellar, conic pillar and cover layers, are found on the mature eg-shell. Silkmoth chorion proteins were detected more than 80 components from a single chorion by two-dimensional electrophoresis. Major protein components of the egg-shell have bee identified on the basis of their isoelectric points and molecular weights, pH 4-6 and 6-30 kd. Several protein components are found entirely or predominantly in th cover layers.

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Effects of 4-Nitroquinoline-1-oxide on Synthetic Patterns of Protein and Glycosaminoglycan during Chick Development (계배 발생중의 단백질 및 당점액질 합성에 대한 4NQO의 영향)

  • Hah, Jae-Chung;Han, Bog-Key
    • The Korean Journal of Zoology
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    • v.26 no.4
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    • pp.283-294
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    • 1983
  • In order to study the change of protein synthesis in developing chick embryo under the influence of carcinogenic substances, we introduced a kind of potent carcinogen, 4-nitroquinoline-1-oxide (4 NQO) to the yolk sac of developing chick embryo. There are changes of synthetic pattern of protein abreast with proceeding of development is each tissue. Protein synthesis in samples which was treated with 4NQO was decreased as compared with untreated control samples. There are tissue specificity in 4NQO effect, showing the apparent decreasing at the 24 hr after drug treatment in the lung and gizzard muscle but not in heart muscle. The results in the present study suggested that 4NQO treated samples was decreased as compared with control samples. From the above data we could know that 4NQO also affected the synthesis of glycosaminoglycan.

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Artemisia argyi H.Lev. & Vaniot Inhibits Matrix metalloproteinases in the Interleukin-1β-stimulated Primary Chondrocytes and Attenuates Osteoarthritis Progression in Mice (황해쑥의 Interleukin-1β 유도 연골세포에서 Matrix metalloproteinase들의 억제효과 및 마우스에서 관절염 진행 감소 효과)

  • Park, Chan Hum;Yang, Chang Yeol;Yang, Siyoung;Yokozawa, Takako;Shin, Yu Su
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.94-94
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    • 2019
  • 황해쑥(Artemisia argyi H.Lev. & Vaniot)은 우수한 항염증 활성을 지닌 것으로 다양하게 보고 되었다. 그러나, 대표적인 염증 질환 중 하나인 골관절염에 미치는 영향은 현재까지 알려져 있지 않다. 따라서, 본 연구에서는 염증 유발 연골 세포 및 골관절염 유발 동물 모델에 미치는 황해쑥 효과에 대해 조사하였다. 첫째, interleukin 1 beta를 투여한 관절 연골 세포에 황해쑥 물 추출물을 투여한 후 metalloporeinase (MMP) -3 및 MMP-13의 발현을 mRNA 및 단백질 수준에서 분석 하였다. 또한, 내측 반월상 연골의 불안정화에 의해 유도 된 골관절염 마우스 모델을 사용하여 황해쑥 물 추출물의 골관절염 억제 효과를 분석 하였다. 세포 실험에서, 본 황해쑥은 MMP-3와 MMP-13의 mRNA 및 단백질 발현을 유의하게 억제하였다. 또한. 황해쑥 물 추출물을 투여한 실험 동물의 관절 조직을 Safranin O 염색을 실시하여 분석한 결과 연골 하 골판 두께의 감소 및 활막 염증 개선 효과가 관찰 되었다. HPLC를 이용한 성분 분석 결과, 황해쑥 물 추출물은 항염증 및 항관절염 활성을 가진 jaceosidin과 eupatilin을 함유하는 것으로 나타났다. 본 연구결과로부터 황해쑥은 골관절염의 치료 또는 예방에 유망한 소재로 개발될 수 있음을 시사한다.

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Roles of Sonic Hedgehog Signaling During Tooth Root and Periodontium Formation (치근 및 치주조직 형성과정 동안 Sonic Hedgehog signaling의 역할)

  • Hwang, Jaewon;Cho, Eui-sic;Yang, Yeonmi
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.2
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    • pp.144-153
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    • 2018
  • The aim of this study was to understand the roles of Sonic Hedgehog (SHH) signaling during tooth root and periodontium formation. In this study, we generated the dental mesenchyme-specific Smoothened (Smo) activated/inactivated mice with the activity of Cre recombinase under the control of osteocalcin promoter. In the Smo activated mutant molar sections at the postnatal 28 days, we found extremely thin root dentin and widened pulp chamber. Picrosirius red staining showed loosely arranged fibers in the periodontal space and decreased cellular cementum with some root resorption. Immunohistochemical staining showed less localization of matrix proteins such as Bsp, Dmp1, Pstn, and Ank in the cementum, periodontal ligament, and/or cementoblast. In the Smo inactivated mutant mouse, there was not any remarkable differences in the localization of these matrix proteins compared with the wild type. These findings suggest that adequate suppressing regulation of SHH signaling is required in the development of tooth root and periodontium.

Synaptic Pattern of NMDA R1 upon the Direction-Selective Retinal Ganglion Cells in Developing Mouse Retina (발생 중 마우스 망막에서 방향특이성 신경절세포의 NMDA R1 수용체의 시냅스 패턴)

  • Lee, Jee-Geon;Kwon, Oh-Ju;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.4
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    • pp.533-540
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    • 2013
  • Purpose: To investigate the synaptic pattern of NMDA glutamate receptor subtype NMDA R1 on the dendritic arbors of ON-OFF direction-selective retinal ganglion cells (DS-RGSs) in developing [(5,10) days postnatal (PN)] mouse retina. Methods: ON-OFF DS-RGCs were injected with Lucifer yellow and the cells were identified by their characteristic morphology. To identify glutamatergic excitatory input from bipolar cell, we used a marker for the membrane traffic motor protein kinesin. Results: We identified DS-RGCs in P5, and P10 mouse retina. The immunofluorescence labeling of NMDA R1 was most prominent in the IPL. Our results showed that their presence upon the entire dendritic arbor of ON-OFF DS-RGCs is without any evidence of asymmetry, which would predict direction selectivity. Conclusions: The glutamatergic input from bipolar cell reveals symmetry pattern in all periods of P5, and P10. The results may suggest that direction selectivity not lies in the specific pattern of NMDA R1 receptors.

Functional Analysis of Expressed Sequence Tags from Hanwoo (Korean Cattle) cDNA Libraries (한우 cDNA 라이브러리에서 발현된 ESTs의 기능분석)

  • Lim, Da-Jeong;Byun, Mi-Jeong;Cho, Yong-Min;Yoon, Du-Hak;Lee, Seung-Hwan;Shin, Youn-Hee;Im, Seok-Ki
    • Journal of Animal Science and Technology
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    • v.51 no.1
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    • pp.1-8
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    • 2009
  • We generated 57,598 expressed sequence tags (ESTs) from 3 cDNA libraries of Hanwooo (Korean Cattle), fat, loin, liver. Liver, intermuscular fat and longissimus dorsi tissues were obtained from a 24-month-old Hanwoo steer immediately after slaughter. cDNA library was constructed according to the oligocapped method. The EST data were clustered and assembled into unique sequences, 4,759 contigs and 7,587 singletons. To carry out functional analysis, Gene Ontology annotation and identification of significant leaf nodes were performed that were detected by searching significant p-values from $2^{nd}$ level GO terms to leaf nodes using Bonferroni correction. We found that 13, 26 and 8 significant leaf nodes are unique in the transcripts according to 3 GO categories, molecular function, biological process and cellular component. Also digital gene expression profiling using the Audic's test was performed and tissue specific genes were detected in the above 3 libraries.

Establishment and characterization of porcine mammary gland epithelial cell line using three dimensional culture system (3차원 배양 시스템을 이용한 돼지 유선 상피 세포 주 특성과 설정)

  • Chung, Hak-Jae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.10
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    • pp.551-558
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    • 2017
  • To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

Moleculay Cloning of the cDNA Encoding the 16 kDa Subunit of V-ATPase in Rat Brain (흰쥐 뇌에서 발현되는 16 kDa Vacuolar (H$^{+}$)-ATPase의 유전자 클로닝)

  • Shin, Song-Woo;Yoo, Min
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.165-170
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    • 2000
  • Vacuolar (H$^{+}$)-ATPase (V-ATPase) is an intracellular protein which consists of multiple subunits. It carries out acidification by pumping protons in the cell. This enzyme has also been found in the synaptic vesicles and may play an important role in the neurotransmission. We cloned cDNA fragments encoding the 16 kDa subunit of V-ATPase from the rat brain by RT-PCR and PCR using total RNA or recombinant phage DNA as templates. They contained the full coding sequences (468 bp) and one nucleotide at 3' region turned out to be different (A to C) when compared to the liver counterpart. However, this polymorphic difference did not cause any significant change in the primary structure of the protein because both GCA and GCC code for alanine. Our study would contribute to the understanding of the function of 16 M)a V-ATPase in the brain and of the mechanisms of neurotransmission.

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Changes of Proteolytic Activity and Amino Acid Composition of the Tissue Extract from Sea Cucumber Entrails during Fermentation with Salt (해삼내장(內臟)젓갈 숙성중(熟成中) 단백질분해효소(蛋白質分解酵素)의 활성(活性)과 아미노산(酸) 조성(組成)의 변화(變化))

  • Lee, Gi Chan;Cho, Deuk Moon;Byun, Dae Seok;Joo, Hyen Kyu;Pyeun, Jae Hyeung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.4
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    • pp.342-349
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    • 1983
  • This study was undertaken to ascertain food and nutritional evaluating data on the processing of fermented sea cucumber (Stichopus japonicus) entrails. In the experiment, the crude proteolytic enzyme from the entrails tissue of raw and fermented sea cucumber during the days of ripening was extracted. The optimal activity condition for the crude enzyme and the compositional changes of amino acid of the protein and free amino acid in the raw and fermented sample were also investigated. 1. Less than three kinds of proteolytic enzymes that each enzyme has optimal activity condition at pH 3.1 $50^{\circ}C$(A-enzyme), pH 5.7 $50^{\circ}C$(B-enzyme) and pH 7.7 $45^{\circ}C$(C-enzyme), respectively were believed to be exist in the entrails tissue of sea cucumber. 2. A-enzyme and C-enzyme were strongly inhibited with the increase of the salt concentration, and B-enzyme was activated at the 1% salt concentration and was inhibited above the 5% salt concentration. 3. The result of the effect of several salt ions on the proteolytic activity showed that A-enzyme was slightly inhibited in the presence of all salt ions added, B-enzyme was activated in the presence of the all salt ions except $Cu^{2+}$ and C-enzyme was activated in the presence of $Ca^{2+}$ and $Mn^{2+}$, and inhibited by $Cu^{2+}$, $Co^{2+}$ and $Mg^{2+}$. 4. When the effects of the ripening days on the proteolytic activity of the crude enzymes were analysed, the activity of the A-enzyme was slightly weakened with the lapse of the fermentation days, whereas the B-enzyme was not influenced by the fermentation days. 5. In the analysis of amino acid composition of the protein of the samples, the 8 days fermented sea cucumber entrails showed the diminution of all kinds of amino acid. Apparently diminished amino acids were arginine, alanine, glutamic acid, glycine, serine, valine, threonine and lysine etc., and methionine, histidine and isoleucine were slightly decreased. 6. In the analysis of free amino acid composition of the 8 days fermented sample, glutamic acid, aspartic acid, leucine and lysine were rich, while histidine, methionine, proline and tyrosine were poor. The most of free amino acids were increased during the fermentation procedure and especially in lysine, histidine, threonine, glutamic acid, methionine, valine and leucine.

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