• Title/Summary/Keyword: Cell Fusion

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Construction of Astaxanthin Overproducing Strain of Phaffia rhodozyma by Protoplast Fusion

  • Koh, Moo-Suk;Kim, Sang-Moon
    • Journal of Microbiology and Biotechnology
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    • v.2 no.1
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    • pp.46-49
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    • 1992
  • The availability of Phaffia rhodozyma as an astaxanthin sources in the aquaculture industry is limited because of the low carotenoid content of natural isolate. In this study, we have used the protoplast fusion technique to construct cell hybrids with an increased content of astaxanthin from P. rhodozyma. Cell hybrids (F307 and F406) obtained were very stable and produced considerably more astaxanthin (> 1 mg/g yeast) than the wild parent. Karyogamy was confirmed by the isolation of recombinants after mitotic segregation of parental auxotrophic genetic markers, the increased amount of chromosomal DNA/cell and the presence of single nucleus/cell.

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Development of Bipolar Plate Stack Type Microbial Fuel Cells

  • Shin, Seung-Hun;Choi, Young-jin;Na, Sun-Hee;Jung, Seun-ho;Kim, Sung-hyun
    • Bulletin of the Korean Chemical Society
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    • v.27 no.2
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    • pp.281-285
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    • 2006
  • Microbial fuel cells (MFC) stacked with bipolar plates have been constructed and their performance was tested. In this design, single fuel cell unit was connected in series by bipolar plates where an anode and a cathode were made in one graphite block. Two types of bipolar plate stacked MFCs were constructed. Both utilized the same glucose oxidation reaction catalyzed by Gram negative bacteria, Proteus vulgaris as a biocatalyst in an anodic compartment, but two different cathodic reactions were employed: One with ferricyanide reduction and the other with oxygen reduction reactions. In both cases, the total voltage was the mathematical sum of individual fuel cells and no degradation in performance was found. Electricity from these MFCs was stored in a supercapacitor to drive external loads such as a motor and electric bulb.

A Study on Proliferation and Phenotypical Stability of Schwann Cell on Keratin/PLGA Film (케라틴이 첨가된 PLGA 필름에서 케라틴 함량별 SC세포의 증식 및 형태유지에 관한 연구)

  • Oh, A-Young;Kim, Soon-Hee;Kim, Yun-Tae;Jeon, Na-Ri;Yang, Jae-Chan;Lee, Sang-Jin;Yoo, James-J.;Van Dyke, Mark;Shin, Hyung-Sik;Rhee, John-M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.2
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    • pp.118-123
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    • 2009
  • Keratin contains regulatory molecules that can enhance neuronal cell activity. We fabricated keratin/ PLGA films using 0, 10, 20, and 50 wt% of keratin using solvent casting method. We measured the contact angle of each film and cell proliferation was assayed by counting the cells attached on the film. Adhered cell morphology was confirmed by scanning electron microscope. RT-PCR was conducted to evaluate the gene expression of NF, NSE, and S-100, the Schwann cell markers. The keratin content of 20 and 50 wt% provided higher wettability than PLGA. The 20 wt% keratin was better in cell adhesion and proliferation of SCs than other keratin/PLGA films. The phenotypic stability of SC was maintained with the keratin content of 10 and 20 wt%.

Discovery of New Fusion Inhibitor Peptides against SARS-CoV-2 by Targeting the Spike S2 Subunit

  • Kandeel, Mahmoud;Yamamoto, Mizuki;Tani, Hideki;Kobayashi, Ayako;Gohda, Jin;Kawaguchi, Yasushi;Park, Byoung Kwon;Kwon, Hyung-Joo;Inoue, Jun-ichiro;Alkattan, Abdallah
    • Biomolecules & Therapeutics
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    • v.29 no.3
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    • pp.282-289
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    • 2021
  • A novel coronavirus, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), caused a worldwide pandemic. Our aim in this study is to produce new fusion inhibitors against SARS-CoV-2, which can be the basis for developing new antiviral drugs. The fusion core comprising the heptad repeat domains (HR1 and HR2) of SARS-CoV-2 spike (S) were used to design the peptides. A total of twelve peptides were generated, comprising a short or truncated 24-mer (peptide #1), a long 36-mer peptide (peptide #2), and ten peptide #2 analogs. In contrast to SARS-CoV, SARS-CoV-2 S-mediated cell-cell fusion cannot be inhibited with a minimal length, 24-mer peptide. Peptide #2 demonstrated potent inhibition of SARS-CoV-2 S-mediated cell-cell fusion at 1 µM concentration. Three peptide #2 analogs showed IC50 values in the low micromolar range (4.7-9.8 µM). Peptide #2 inhibited the SARS-CoV-2 pseudovirus assay at IC50=1.49 µM. Given their potent inhibition of viral activity and safety and lack of cytotoxicity, these peptides provide an attractive avenue for the development of new prophylactic and therapeutic agents against SARS-CoV-2.

Novel AGLP-1 albumin fusion protein as a long-lasting agent for type 2 diabetes

  • Kim, Yong-Mo;Lee, Sang Mee;Chung, Hye-Shin
    • BMB Reports
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    • v.46 no.12
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    • pp.606-610
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    • 2013
  • Glucagon like peptide-1 (GLP-1) regulates glucose mediated-insulin secretion, nutrient accumulation, and ${\beta}$-cell growth. Despite the potential therapeutic usage for type 2 diabetes (T2D), GLP-1 has a short half-life in vivo ($t_{1/2}$ <2 min). In an attempt to prolong half-life, GLP-1 fusion proteins were genetically engineered: GLP-1 human serum albumin fusion (GLP-1/HSA), AGLP-1/HSA which has an additional alanine at the N-terminus of GLP-1, and AGLP-1-L/HSA, in which a peptide linker is inserted between AGLP-1 and HSA. Recombinant fusion proteins secreted from the Chinese Hamster Ovary-K1 (CHO-K1) cell line were purified with high purity (>96%). AGLP-1 fusion protein was resistant against the dipeptidyl peptidase-IV (DPP-IV). The fusion proteins activated cAMP-mediated signaling in rat insulinoma INS-1 cells. Furthermore, a C57BL/6N mice pharmacodynamics study exhibited that AGLP-1-L/HSA effectively reduced blood glucose level compared to AGLP-1/HSA.

The Mechanism of Membrane Fusion During the Infection of HIV

  • Yu Yeon Gyu
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.97-101
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    • 2001
  • The fusion between viral envelope and target cell membrane is a central step of viral infection, and the fusion proteins located at viral envelope mediate such process. Gp41 of HIV is one of the fusion proteins whose structure and mechanism of membrane fusion had been extensively studied. Functionally important motives of gp41 are the N-terminus fusion peptide, the coiled-coil and the membrane proximal C-peptide regions. The role of these regions during the fusion process had been thoroughly examined. Specially, insertion of the fusion peptide into membrane and conformational change of the coiled-coil and C-peptide regions are assumed to be critical for the fusion mechanism. In addition, the coiled-coil region has been shown to interact with membrane, and the C-peptide region regulates the interaction in a dose dependent manner. Furthermore, fusion defective mutations of the coiled-coil region dramatically changed its binding affinity to membrane. These results suggested that the membrane binding property of the coiled-coil region is important for the fusion activity of gp41, and such property could be modulated by the interaction with the C-peptide region.

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Studies on the Fusion Mechanism of the Cell (1) (細胞의 融合機作에 관한 硏究(1))

  • Kang, Man-Sik;Seunhyon Choe;Wookeun Song
    • The Korean Journal of Zoology
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    • v.26 no.4
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    • pp.235-251
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    • 1983
  • Several approaches have been made to access the mechanism of fusion in chick myoblast in vitro. Lactoperoxidase-catalyzed iodination was applied to labell cell surface proteins during myogenesis. Quantitative as well as qualitative changes were observed in $^131$I surface components of prefusion and postfusion cells. Two proteins with a molecular weight of 165K and 93K daltons were observed to appear at the onset of fusion as compared to prefusion stage. At the same time, 245K dalton protein decreased whereas the low molecular weight proteins increased consistently. The decrease of high molecular weight proteins appears to be associated with the cell cycle of myoblast during differentiation. The increased appearance of low molecular weight proteins might be due to the proteolytic cleavage of the high molecular weight proteins. Examination of intracellulr cAMP levels during fusion has revealed that a large but transient increase in cAMP occurs before the onset of fusion. This result suggests a causal relationship between the increase of cAMP and the onset of fusion, and further, that differentiating myoblasts are synthronized to a high degree. During the course of myoblast differentiation, at least four lowe molecular weight proteins, which different from major surface proteins iodinated, were identifiable in the culture medium. These proteins could be ascribed to be released from the membrane by proteolytic cleavage of surface proteins in the course of myoblast fusion. The significance of cell surface alterations and the released proteins during the fusion, the involvement of cAMP in the onset of fusion and the possibility that fusion is promoted by external factor(s) are discussed.

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Nuclear Transplantation of Bovine IVF Embryos by Cell Cycle Control of Recipient Cytoplasm (수핵란 세포질의 세포주기 조절에 의한 소 체외수정란의 핵이식)

  • 정희태;임석기;박춘근;양부근;김정익
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.307-313
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    • 1996
  • This study was conducted to investigate the effect of S-phase synthronized nuclear transfer on the development of nuclear transplant bovine embryos. A blastomere derived from the 16~32 cell stage bovine embryos was transferred into an enucleated metaphase II(MII) oocytes or activated S-phase eggs. From the MII-phase and S-phase nuclear transfer, 6.3%(4/63) and 13.8%(9/65) of nuclear transplant embryos developed to the blastocyst stage, respectively. In the S-phase nuclear transfer, maximal proportion of embryos developed to the blastocyst stage(16.6%) was obtained after the recipient cell was activated 8 h prior to receving a donor nucleus. MII-phase nuclear transplant embryos showed the PCC state of their nuclear at 1.5~2 h after fusion, whereas, S-phase nuclear transplant embryos did not undergo PCC. The result of this study suggests that if blastomeres of unknown cell-cycle-stage are used, S-phase nuclear transplantation through the activation of enucleated oocytes prior to fusion enhances development of nuclear transplant embryos. This result also suggests that the interval time from oocyte activation to cell fusion may affect the development of nuclear transplant embryos.

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Development of Diagnostic System to Black Queen Cell Virus(BQCV) Using Multi-point Detection (Multi-point PCR법을 이용한 Black Queen Cell Virus (BQCV) 검출법 개발)

  • Kim, Somin;Kim, Byounghee;Kim, Moonjung;Kim, Jungmin;Truong, A Tai;Kim, Seonmi;Yoon, Byoungsu
    • Journal of Apiculture
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    • v.34 no.1
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    • pp.39-46
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    • 2019
  • BQCV multi-point PCR was developed as a rapid multiplex detection method for BQCV, one of the viral pathogens of honeybees. It could detect BQCV specific genes qualitative as well as quantitative detection based on ultra-rapid PCR. Three primer pairs (RNA dependent RNA polymerase, capsid protein, 3C like protease) were specifically designed for accurate the detection and were optimized for minimizing the detection time and increasing the sensitivity. Our advanced diagnostic system have the accuracy by lowering the concern about the variation in the BQCV detection site. In addition, it should be an opportunity to identify mutations that are mixed with other viruses.