• Title/Summary/Keyword: 세포융합기술

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Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A (보툴리눔 신경독소 A를 중화하는 재조합 항체의 제조와 특성 분석)

  • Park, Hong-Gyu;Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.1
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    • pp.295-301
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    • 2017
  • Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

A Study of Literary Therapy on the Rated Sijo as a Conductor that Works the Motherboard of Mind (마음의 메인보드를 작동시키는 전도체로서의 정격 시조에 관한 문학치료 연구)

  • Park, In-kwa
    • The Journal of the Convergence on Culture Technology
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    • v.2 no.4
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    • pp.31-40
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    • 2016
  • The hardware of the human body is given the life force by the sentence which is the physiological software that the program for cell activation by the electrical signal enters. The aim of this study is to create a better therapeutic environment for the human body that groaned with errors in the physiological and cognitive systems that are transmitted to neurons and neurons. The sentence program of the rated sijo, which is the software of the human body which has the function as a conductor to connect the emotions of joy, anger, sadness, and enthusiasm to the human mental system, can be connected to the neuron system of the human body, we tried to identify the principle of operating the motherboard of mind in humanities. Once these principles are identified, we can figure out how to minimize side effects and lead the body to a therapeutic program. The research found that there is a strong energy source that can operate the motherboard of the heart very quickly in the rated Sijo. This is because it is confirmed that new coding and re-coding of a number of rated sijo, or a new syllable of one syllable followed by the original syllable of the original syllable, are formed quickly and therapeutically.This has led to the possibility of literary therapy for mankind to upgrade the human psychic system in abundance through the function of the interaction between the sentence as a conductor that is synaptically connected to the human body and the mainboard of the mind attached to the human body without side effects in the future.

Studies on the tolerance of Halophyte Arabis stelleri under heavy metals and Salt stress condition (염생식물 섬갯장대(Arabis stelleri var. japonica)의 중금속 및 고염 농도 스트레스 상태에서 내성 연구)

  • Kim, Donggiun
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.4
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    • pp.373-378
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    • 2019
  • In the marine area, the salt concentration in the soil increases, and the inland heavy metal pollution increases the damage of plants. In the inland industrial development area, researches on the genetic resources of plants together with the heavy metal accumulation of Co, Ni, Zn, and so on are required. Both of these problems have caused scientists to work hard to find plants that are likely to cause stress in plant roots. In this study, seeds of Arabis stelleri var. japonica collected near the shore were used for germination. The growth and development and tolerance of both Arabis and Arabidopsis seeds were investigated under laboratory culture conditions. As a result, Arabis showed resistance about 3 times in 250 mM nickle and cobalt, and more than 4 times in 1 mM zinc when compared to Arabidopsis. The tolerance of Arabis to Na salts increased by 20% or more at 50 mM concentration and Arabis was resistant to heavy metals and salt concentration. The accumulation of Na ions in the body was measured as a preparation for studying the intracellular mechanism. As a result, it showed a further decrease in resistance to ground water roots. It is considered that the activity of the exporting gene is important rather than the mechanism of accumulation.

Biolagical Activity on Extracts of Japanese Anise(Illicium Anisatum L.) Leaves and Twigs (붓순나무 잎과 가지의 추출물에 대한 생리활성 평가)

  • Shinn, Seong-Whan
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.3
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    • pp.311-316
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    • 2019
  • Japanese anise (Illicium anisatum L.) leaves and twigs were extracted with 50 % aqueous acetone three times. After filtration, the extracts were fractionated with n-hexane, chloroform, ethyl acetate and $H_2O$, and then freeze dried after condensation. Then antioxidation and antiviral activity were evaluated on each fractions. In the antioxidative activities, the results indicated high activity in the EtOAc soluble fraction of the leaves and the EtOAc and $H_2O$ soluble fractions of the twigs. It showed much higher antioxidative value compare to the controls, BHT and ${\alpha}$-tocopherol. In the antiviral activities, the all fractions were negative effects in HRV 1B and EV 71, but good in Influenza PR8. The activities of the crude extracts of the leaves and twigs showed more than 80% activity at the concentration of $10{\mu}g/mL$ and $50{\mu}g/mL$, respectively, and the activities of the EtOAc and $H_2O$ soluble fractions were close to 80%. Based on the above results, the extracts of Japanese anise may be applied for one of the natural biomass sources that can be used as an antioxidant and an antiviral substance.

On a Cleaning of COVID-19 Prevention Masks with Electrolytic Decomposition Water (전기분해수로 코로나방역용 마스크의 세정에 관한연구)

  • Tian, Zhixing;Bae, Myung-Jin
    • The Journal of the Convergence on Culture Technology
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    • v.8 no.1
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    • pp.591-596
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    • 2022
  • Various COVID-19 quarantine guidelines and measures are being taken by country at the WHO, but the number of confirmed cases has not decreased significantly. In order to prevent the inflow and outflow of COVID-19 through individual droplets, it is mandatory to wear a mask anytime, anywhere. However, as virus bacteria entering the mask amplify, it pollutes the mask and causes a disgusting smell. In this paper, a new method of preventing the spread of COVID-19 was proposed by sterilizing the mask with a dental gait spray introduced into the mask that has been used for a long time. Dental gargle water is usually produced by electrolysis of tap water, and the unstable ion water (HOCl) dissolved in water penetrates the cell barrier of various viruses and fails to act in its nucleus, causing water to self-purify. As a result of the experiment, when the mask used for a long time was washed with gargle water spray, the washed mask was dried after 10 minutes, and the smell of virus droplets or saliva almost disappeared. In particular, as a result of MOS testing the fit of the subjects who participated in the mask cleaning, it was excellent at 4.4 on average. Therefore, the mask was disposable, but if the spray was washed in the proposed method more than twice a day, the mask could be used in a comfortable state for more than a week.

Development of Anti-Aging Products (Anti-Wrinkle) like Epidermal Growth Factor(EGF) Materials using Supercritical Heat-Treated Extract Radish (초임계 열처리된 무 성분을 이용한 상피세포성장인자(EGF) 유사소재 개발 및 광노화(주름개선) 효과)

  • Kim, Hyun Kyoung
    • The Journal of the Convergence on Culture Technology
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    • v.4 no.3
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    • pp.197-207
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    • 2018
  • The radish skin and radish greens (mucheong) are an edible part of the radish. But they are removed before eating the radish and used as a byproduct or an animal feed material because of their tough and rough texture. This study was conducted to investigate the effect of supercritical heat-treated radish-extract on UV-induced Hos: HRM-2 wrinkled mouse animal model on anti-aging wrinkles. Supercritical heat-treated radish-extract was applied on the back of seven-weeks old HRM-2 mice. The effect of HRE on skin thickness, elasticity and wrinkle formation of the mice was observed by using UVB lamp to induce melanogenesis and wrinkle formation. As the result, increased depth of wrinkles was observed in the negative control group in comparison to the normal group. In contrast, decreased depth of wrinkles was observed in the radish-extract-free group compared to the negative control group. In the study of the effect of radish-extract on wrinkle-formation related gene expression and protein what protein expression, MMP-2 and MMP-2 gene expression significantly increased in the negative control group compared to the normal group. The gene expression reduced independence to the mass of radish-extract treated. Similar to quantitative results of mRNA expression, the expression of MMP-2 protein increased as a result of UVB-irradiation. The MMP-2 expression was inhibited in dependence to the mass of radish-extract treated. In conclusion, the supercritical heat-treated radish-extract has an effect on improving skin wrinkles not only when it is applied to the skin but also when orally ingested. Thus, it can be effectively used as a composition to health functional products. Thereafter, we can also conclude that radish, a food that does not show any side-effects even upon long-term intake, can reduce wrinkle formation as well as improve skin elasticity when taken regularly for a long period.

Production of a Monoclonal Antibody to Human $\alpha$-Fetopotein and Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for Human $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein에 대한 모노클로날 항체의 제조 및 모노클로날 항체를 이용한 효소면역분석법의 개발)

  • Michung Yoon;Hyun-Hee Lee;Youngwon Lee
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.1-10
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    • 1999
  • This study was attempted to generate a monoclonal antibody against human $\alpha$-fetoprotein (AFP) and to produce an immunoassay, recognizing AFP in plasma and amniotic fluid. AFP was purified from human amniotic fluid and used to immunize mice. Spleens were taken from the mice and the cells were fused with mouse myeloma cells (Sp2/0-Ag-14) for the production of monoclonal antibodies by employing the hybridoma technology. As a result, a hybridoma cell line producing anti-AFP monoclonal antibody was cloned out and designated as MabF22. From isotyping analysis, it was found that monoclonal antibody MabF22 was IgG type with IgG1 heavy chain and k light chain. The binding specificity of MabF22 was analyzed by immunoblotting as well as by ELISA. MabF22 was highly specific, reacting with only AFP-containing samples. The binding affinity was determined by ELISA (free-capture mode) and Scatchard analysis. As a result, the value of Kd was 0.8$\times$10$^{-10}$M. The validity of the MabF22 for AFP assay was examined by two kinds of ELISAs, i.e., non-competitive and competitive ELISA. Both assays revealed that MabF22 reacted well with AFP in sample in a concentration-dependent manner. Standard curve and antibody titration curve were obtained by using purified AFP and MabF22. These results indicate that the monoclonal antibody produced in this study would be useful not only for research purposes but also for further development of immune-diagnostic kit for the measurement of AEP concentration.

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Expression of the lux Genes in Escherichia coli for the Basis of Development of Biosensor (바이오센서 개발을 위한 발광 유전자의 대장균에서의 발현 시스템 분석)

  • Cho, Mi-Mi;Kim, Young-Doo;Kang, Kyung-Sook;Kim, Sook-Kyung;Yang, In-Chul;Park, Sang-Ryoul;Lee, Chan-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.419-424
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    • 2009
  • To provide the basis of biosensor based on the lux genes from bioluminescent bacteria of Photobacterium leiognathi and Vibrio harveyi, we test the expression of lux genes in several strains of Escherichia coli. The expression of the recombinant plasmid of PlXba.pT7-3, containing all lux genes requiring for light emission without adding substrate, in E. coli 43R was so strong to see the blue-green light in single colony as well as in the alginate immobilized cell. In addition, the light intensity was decreased by adding heavy metal ion such as cadmium and zinc ions. These result raise the possibility that a biosensor can be developed using the lux genes system.

Antiaging Activity of Mixed Extracts from Korean Medicinal Herbs on HS68 Skin Fibroblast (한약재복합 추출물의 인간피부섬유아세포 HS68에 대한 항노화 효과)

  • Shin, Dong-Chul;Kim, Gwui-Cheol;Song, Si-Young;Kim, Hee-Jin;Yang, Jae-Chan;Lee, Yong-Hwa;Kim, Bo-Ae
    • The Korea Journal of Herbology
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    • v.29 no.2
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    • pp.39-45
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    • 2014
  • Objectives : The purpose of this study was to investigate antiaging and antioxidant effects on cultured human skin fibroblast with 80% ethanol extracts of plants including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Methods : An ethanol extract of three medicinal plants including stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Extracts were assessed to determine the mechanism of antioxidant and antiaging activities. Antioxidant activity of extract was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. These extracts were tested for cell viability on HS68 skin fibroblast by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. We investigated the effects of Ultraviolet-B irradiation on cytotoxicity, type 1 collagen, elastin level and oxidative damage in cultured human skin fibroblast (HS68). Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Results : The extracts obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The extracts of complex herbal medicine showed low cytotoxicity as more than 100% cell viability in 100ppm/ml concentration. HS68 fibroblasts were survived 70% at 120 $mJ/cm^2$ UVB irradiation and treated tumor necrosis factor (TNF)-alpha. The levels of aging factors and cytotoxicity were decreased by ethanol extract of complex herbal medicine. Conclusions : These results suggest that ethanol extracts of complex medicinal plants of including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus may have value as the potential antioxidant and antiaging medicinal plant.

Artificial Mutation for Silkworm Molecular Breeding Using Gene Scissors (유전자 가위의 이용과 누에 분자 육종을 위한 인위적 돌연변이 유발)

  • Hong, Jeong Won;Jeong, Chan Young;Yu, Jeong Hee;Kim, Su-Bae;Kang, Sang Kuk;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Park, Jong Woo
    • Journal of Life Science
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    • v.30 no.8
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    • pp.701-707
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    • 2020
  • Gene editing technology using the clustered regularly interspaced short palindromic repeat (CRISPR) and the CRISPR associated protein (Cas)9 has been highly anticipated in developing breeding techniques. In this study, we discuss gene scissors as a tool for silkworm molecular breeding through analysis of Bombyx mori Kynurenine 3-Monooxygenase (BmKMO) gene editing using the CRISPR/Cas9 system and analysis of generational transmission through mutagenesis and selective crossing. The nucleotide sequence of the BmKMO gene was analyzed, and three guide RNAs (gRNAs) were prepared. Each synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA and Cas9 complexes were subsequently microinjected into the silkworm embryos; the hatching rate was 18% and the incidence of mutation was 60%. The gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed. In homozygotes generated by self-crossing, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and could be an effective way of shortening the time required.