• Title/Summary/Keyword: C1-2 fusion

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Preparation of Korean Traditional Alcoholic Beverage (Yakju) by a Protoplast Fusion Yeast Strain Utilizing Starch and its Quality Characteristics (전분분해 효모융합체를 이용한 전통 발효주의 제조와 품질특성)

  • Ju, Min-No;Hong, Sung-Wook;Kim, Kwan-Tae;Yum, Sung-Kwan;Kim, Gye-Won;Chung, Kun-Sub
    • Korean Journal of Food Science and Technology
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    • v.41 no.5
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    • pp.541-546
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    • 2009
  • The objectives of this study were to evaluate the physico-chemical and sensory characteristics of a Korean traditional alcoholic beverage (yakju) prepared using different nuruk (Korean-style koji) concentrations and yeasts such as the fusant FA776 and Saccharomyces cerevisiae KOY-1, respectively. The fusant FA776, which has alcohol-fermenting and starch-utilizing properties, was formed by Saccharomyces cerevisiae KOY-1 and Saccharomyces diastaticus KCTC1804. The fermentation trial was conducted in a 5 L lab-scale jar at $25^{\circ}C$. The maximum alcohol production of the K-100 and F-50 reached levels of 135.0 mg/mL and 119.4 mg/mL, respectively. The pH values were in a range of 4.3-4.5. Total acidity was in a range of 0.47-0.60%. Organic acids and amino acids were analyzed in order to evaluate variations in its composition and content via HPLC analysis. Organic acids including lactic acid, citric acid, malic acid, and pyruvic acid, and 16 kinds of amino acids, including aspartic acid, were detected in all treatments. K-100 showed the highest amino acid contents, whereas F-50 exhibited the lowest amino acid contents. Volatile flavor components such as phenylethyl alcohol, isoamyl alcohol, 2-methylthiophane, isobutyl alcohol, and ethyl succinate were detected as a major component in all treatments, as determined via gas chromatography. The results of our sensory evaluation demonstrated that Yakju fermented by the FA776 fusant yielded more favorable results than S. cerevisiae KOY-1.

Cell Migration and Wound Healing Activities of Recombinant Thymosin β-4 Expressed in Escherichia coli (재조합 Thymosin β-4의 세포이동능과 상처치유능)

  • Hong, Kyo-Chang;Choi, Yung Hyun;Kim, Gun-Do;Cha, Hee-Jae;Jeon, Sung-Jong;Nam, Soo-Wan
    • Journal of Life Science
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    • v.32 no.2
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    • pp.135-141
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    • 2022
  • Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

Study on ICT convergence in Lentinula edodes (Shiitake) cultivation system using Automated container (컨테이너형 수출용 버섯식물공장시스템설계 및 표고버섯 생산 연구)

  • Jo, Woo-Sik;Lee, Sung-Hak;Park, Woo-Ram;Shin, Seung-Ho;Park, Chang-Min;Oh, Ji-Hyun;Park, Who-Won
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.264-268
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    • 2017
  • In the 21st century, information and communication technology (ICT) worldwide presents a new vision for agriculture. Time and place, as well as the high-tech industry, to overcome barriers to the fusion of the so-called "smart agriculture," are changing the agricultural landscape. Core container production in precision agriculture for mushroom cultivation, optimal temperature, humidity, irradiation, self-regulation of factors such as carbon dioxide, and environment for mushroom cultivation were adopted. Lentinula edodes (shiitake) is an edible mushroom native to East Asia, cultivated and consumed in many Asian countries. It is considered to be medicinal in certain practices of traditional medicine. We used different controlled light sources (Blue-Red-White-combined LED, blue LED, red LED, and fluorescent light) with different LED radiation intensities (1.5, 10.5, and $20.5{\mu}mol/m^2s$ for LEDs) to compare growth and development. Mushrooms were treated with light in a 12-hour-on/12-hour-off cycle, and maintained in a controlled room at $19{\sim}21^{\circ}C$, with 80~90% humidity, and an atmospheric $CO_2$ concentration of 1,000 ppm for 30 days. Growth and development differed with the LED source color and LED radiation intensity. Growth and development were the highest at $10.5{\mu}mol/m^2s$ of blue LED light. After harvesting the fruit bodies, we measured their weight and length, thickness of pileus and stipe, chromaticity, and hardness. The $10.5{\mu}mol/m^2s$ blue-LED-irradiated group showed the best harvest results with an average individual weight of 39.82 g and length of 64.03 mm, pileus thickness of 30.85 mm and pileus length of 43.22 mm, and stipe thickness of 16.96 mm with fine chromaticity and hardness. These results showed that blue LED light at $10.5{\mu}mol/m^2s$ s exerted the best effect on the growth and development of L. edodes (shiitake) mushroom in the ICT-system container-type environment.

Enhancement of PLED lifetime using thin film passivation with amorphous Mg-Zn-F

  • Kang, Byoung-Ho;Kim, Do-Eok;Kim, Jae-Hyun;Seo, Jun-Seon;Kim, Hak-Rin;Lee, Hyeong-Rag;Kwon, Dae-Hyuk;Kang, Shin-Won
    • Journal of Information Display
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    • v.11 no.1
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    • pp.8-11
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    • 2010
  • In this study, a new thin films passivation technique using Zn with high electronegativity and $MgF_2$, a fluorine material with better optical transmittance than the sealing film materials that have thus far been reported was proposed. Targets with various ratios of $MgF_2$ to Zn (5:5, 4:6 and 3:7) were fabricated to control the amount of Zn in the passivation films. The Mg-Zn-F films were deposited onto the substrates and Zn was located in the gap between the lattices of $MgF_2$ without chemical metathesis in the Mg-Zn-F films. The thickness and optical transmittance of the deposited passivation films were approximately 200 nm and 80%, respectively. It was confirmed via electron dispersive spectroscopy (EDS) analysis that the Zn content of the film that was sputtered using a 4:6 ratio target was 9.84 wt%. The Zn contents of the films made from the 5:5 and 3:7 ratio targets were 2.07 and 5.01 wt%, respectively. The water vapor transmission rate (WVTR) was determined to be $38^{\circ}C$, RH 90-100%. The WVTR of the Mg-Zn-F film that was deposited with a 4:6 ratio target nearly reached the limit of the equipment, $1\times10^{-3}\;gm^2{\cdot}day$. As the Zn portion increased, the packing density also increased, and it was found that the passivation films effectively prevented the permeation by either oxygen or water vapor. To measure the characteristics of gas barrier, the film was applied to the emitting device to evaluate their lifetime. The lifetime of the applied device with passivation was increased to 25 times that of the PLED device, which was non-passivated.

Histopathological observation of the gill of the crucian carp, Carassius auratus by the leech, Limnotrachelobdella sinensis (거머리, Limnotrachelobdella sinensis의 기생으로 인한 붕어, Carassius auratus 아가미의 조직병리학적 관찰)

  • Park, Myoung-Ae;Kim, Seok-Ryel;Kim, Myoung-Sug;Kim, Jeong-Ho;Park, Jung-Jun
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.399-407
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    • 2010
  • On the inner side of each operculum of the crucian carp, Carassius auratus (n=10), the leech, Limnotrachelobdella sinensis of 1-4 individuals were parasitic. The leeches had approximately 41.0 mm in total length and 11 mm in width. These body was composed with anterior sucker, neck, trunk and posterior sucker and average length was 2.3 mm, 7.2 mm, 23.3 mm and 8.7 mm respectively. To both sides of the trunk lateral vesicle of 11 pair existed. When observed by SEM, anterior sucker was hemisphere shape and the mouth where proboscis comes out existed with the its center. Proboscis was connected the esophagus directly. Under light microscopy, bloodsucking gill of C. auratus showed lamella fusion, hypertrophy the epithelial cell of the filament and lamella, increased mucocytes and congested capillaries. On the other hand, necrotic and hydropic degeneration epithelial cell of the lamella, and infiltration of the macrophages from some individuals were suggested the secondary infection with the bacteria or virus after bloodsucking activity of the leech.

Dlx3 Plays a Role as a Positive Regulator of Osteoclast Differentiation

  • Cha, Ji-Hun;Ryoo, Hyun-Mo;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.32 no.3
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    • pp.85-91
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    • 2007
  • Dlx3 is a homeodomain protein and is known to playa role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM # 190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. Although the observed defects of TDO syndrome involves bone, little is known about the role of Dlx3 in bone remodeling process. In this study, we examined the effect of wild type DLX3 (wtDlx3) expression on osteoclast differentiation and compared it with that of 4-BP DEL DLX3 (TDO mtDlx3). To examine whether Dlx3 is expressed during RANKL-induced osteoclast differentiation, RAW264.7 cells were cultured in the presence of receptor activator of nuclear factor-B ligand (RANKL). Dlx3 protein level increased slightly after RANKL treatment for 1 day and peaked when the fusion of prefusion osteoclasts actively progressed. When wtDlx3 and TDO mtDlx3 were overexpressed in RAW264.7 cells, they enhanced RANKL-induced osteoclastogenesis and the expression of osteoclast differentiation marker genes such as calcitonin receptor, vitronectin receptor and cathepsin K. Since osteoclast differentiation is critically regulated by the balance between RANKL and osteoprotegerin (OPG), we examined the effect of Dlx3 overexpression on expression of RANKL and OPG in C2C12 cells in the presence of bone morphogenetic protein 2. Overexpression of wtDlx3 enhanced RANKL mRNA expression while slightly suppressed OPG expression. However, TDO mtDlx3 did not exert significant effects. This result suggests that inability of TDO mtDlx3 to regulate expression of RANKL and OPG may contribute to increased bone density in TDO syndrome patients. Taken together, it is suggested that Dlx3 playa role as a positive regulator of osteoclast differentiation via up-regulation of osteoclast differentiation-associated genes in osteoclasts, as well as via increasing the ratio of RANKL to OPG in osteoblastic cells.

A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Quercetin Inhibits the 5-Hydroxytryptamine Type 3 Receptor-mediated Ion Current by Interacting with Pre-Transmembrane Domain I

  • Lee, Byung-Hwan;Jung, Sang-Min;Lee, Jun-Ho;Kim, Jong-Hoon;Yoon, In-Soo;Lee, Joon-Hee;Choi, Sun-Hye;Lee, Sang-Mok;Chang, Choon-Gon;Kim, Hyung-Chun;Han, YeSun;Paik, Hyun-Dong;Kim, Yangmee;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.20 no.1
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    • pp.69-73
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    • 2005
  • The flavonoid, quercetin, is a low molecular weight substance found in apple, tomato and other fruit. Besides its antioxidative effect, quercetin, like other flavonoids, has a wide range of neuropharmacological actions including analgesia, and motility, sleep, anticonvulsant, sedative and anxiolytic effects. In the present study, we investigated its effect on mouse 5-hydroxytryptamine type 3 ($5-HT_{3A}$) receptor channel activity, which is involved in pain transmission, analgesia, vomiting, and mood disorders. The $5-HT_{3A}$ receptor was expressed in Xenopus oocytes, and the current was measured with the two-electrode voltage clamp technique. In oocytes injected with $5-HT_{3A}$ receptor cRNA, quercetin inhibited the 5-HT-induced inward peak current ($I_{5-HT}$) with an $IC_{50}$ of $64.7{\pm}2.2{\mu}M$. Inhibition was competitive and voltage-independent. Point mutations of pre-transmembrane domain 1 (pre-TM1) such as R222T and R222A, but not R222D, R222E and R222K, abolished inhibition, indicating that quercetin interacts with the pre-TM1 of the $5-HT_{3A}$ receptor.

Role of CopA to Regulate repABC Gene Expression on the Transcriptional Level (전사 수준에서 repABC 유전자 발현을 조절하는 CopA 단백질의 역할)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;In Gyu Baek;Kyu Ho Bang
    • Journal of Life Science
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    • v.34 no.2
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    • pp.86-93
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    • 2024
  • Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.