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Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

Co-occurrence of Matsumuraeses falcana and M. phaseoli (Lepidoptera: Tortricidae) in Soybean Fields, and Polymorphism of Cytochrome c Oxidase Subunit 1 Gene Nucleotide (콩 포장에서 어리팥나방과 팥나방(나비목: 잎말이나방과)의 동시 발생과 시토크롬 c 산화효소 1 유전자 염기서열의 다형성)

  • Jin Kyo, Jung;Eun Young, Kim;Taeman, Han
    • Korean journal of applied entomology
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    • v.61 no.4
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    • pp.641-649
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    • 2022
  • Leaf-rolling moths were collected from soybean fields and identified as Matsumuraeses falcana and Matsumuraeses phaseoli by comparison with laboratory-reared species based on the nucleotide sequence (658 bp) of the mitochondrial cytochrome c oxidase 1 subunit gene (COX1). Ten haplotypes with 0.15-0.46% genetic distance from each other in COX1 were found in 47 samples of M. falcana, in which haplotype A was dominant (approximately 70%). Only one type of COX1 was found in 30 samples of M. phaseoli, and its sequence showed 4.11-4.61% genetic distance from those of M. falcana. Amino acid sequences translated from COX1 were identical in all samples of both species, and they showed synonymous substitutions. Larvae of both species caused damage to soybean leaves and pods and co-occurred simultaneously in the field. The average density of M. falcana was 1.5 times higher than that of M. phaseoli. The results clearly indicate that soybean was the host plant for both species. In addition, Elodia flavipalpis (Diptera: Tachinidae) was found to be a larval parasitoid of Matsumuraeses sp. through identification of the COX1 gene.

High-level Expression and Characterization of the Human Interleukin-10 in the Milk of Transgenic Mice

  • Zneng, Z. Y.;B. H. Sohn;K. B. Oh;W. J. Shin;Y. M. Han;Lee, K. K.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.46-46
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    • 2003
  • Interleukin-10 (IL-10) is a homodimeric protein with a wide spectrum of anti-inflammatory and immune activities. It inhibits cytokine production and expression of immune surface molecules in various cell types. The transgenic mice carrying the human IL-10 gene in conjunction with the bovine $\beta$-casein promoter produced the human IL-10 in milk during lactation. Transgenic mice were generated using a standard method as described previously. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues with a primer set. In this study, stability of germ line transmission and expression of IL-10 gene integrated into host chromosome were monitored up to generation F15 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to F15 mice showed similar transmission rates (66.0$\pm$20.13%, 61.5$\pm$16.66%, 41.1$\pm$8.40%, 40.7$\pm$20.34%, 61.3$\pm$10.75%, 49.2$\pm$18.82%, and 43.8$\pm$25.91%, respectively), implying that the IL-10 gene can be transmitted stably up to long term generation in the transgenic mice. For ELISA analysis, IL-10 expression levels were determined with an hIL-10 ELISA and a mIL-10 ELISA kit in accordance with the supplier's protocol. Expression levels of human IL-10 from milk of generation F9 to F13 mice were 3.6$\pm$1.20 mg/ml, 4.2$\pm$0.93 mg/ml, 5.7$\pm$1.46 mg/ml, 6.3$\pm$3.46 mg/ml, and 6.8$\pm$4.52 mg/ml, respectively. These expression levels are higher than in generation F1 (1.6 mg/ml) mice. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations, although there was a little fluctuation in the transmission frequency and expression level between the generations.

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Phylogenetic Analysis of Cucurbit Chlorotic Yellows Virus from Melon in 2020 in Chungbuk, Korea (2020년 충북지역 멜론에서 발생한 Cucurbit Chlorotic Yellows Virus의 계통분석)

  • Taemin Jin;Hae-Ryun Kwak;Hong-Soo Choi;Byeongjin Cha;Jong-Woo Han;Mikyeong Kim
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.52-59
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    • 2023
  • Cucurbit chlorotic yellows virus (CCYV) is a plant virus that causes damage to cucurbit crops such as watermelon and cucumber, and is transmitted by an insect vector known as the whitefly. Since CCYV was first detected on cucumber in Chungbuk in 2018, it has been reported in other areas including Gyeongsang in Korea. In 2020, we performed field surveys of yellowing diseases in the greenhouses growing melon and watermelon in Chungbuk (Jincheon and Eumseong). Reverse transcription-polymerase chain reaction analysis of 79 collected samples including melon, watermelon, and weeds resulted in detection of CCYV in 4 samples: Three samples were singly infected with CCYV and one samples was mixed infected with CCYV, Cucurbit aphid borne yellows virus, and Watermelon mosaic virus. The complete genome sequences of the four collected CCYV melon isolates (ES 1-ES 4) were determined and genetically compared with those of previously reported CCYV isolates retrieved from GenBank. Phylogenetic analyses of RNA 1 and 2 sequences revealed that four ES isolates were clustered in one group and closely related to the CCYV isolates from China. The analysis also revealed very low genetic diversity among the CCYV ES isolates. In general, CCYV isolates showed little genetic diversity, regardless of host or geographic origins. CCYV has the potential to pose a serious threat to melon, watermelon, and cucumber production in Korea. Further studies are needed to examine the pathogenicity and transmissibility of CCYV in weeds and other cucurbits including watermelon.

First Report of Fire Blight Caused by Erwinia amylovora on Korean Mountain Ash (Sorbus alnifolia) in Korea (Erwinia amylovora에 의한 팥배나무 화상병 발생 보고)

  • Yeon-Jeong Lim;Hyeonseok Oh;Mi-Hyun Lee;Eunjung Roh;Hyeonheui Ham;Dong Suk Park;Duck Hwan Park;Yong Hwan Lee
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.79-81
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    • 2023
  • During the nationwide survey of fire blight, the typical shoot blight symptoms were found on Korean mountain ash (Sorbus alnifolia) which was located near an orchard that produced fire blight on pear trees in Eumseong, Korea, May 2021. To identify the causal agent, we progressed isolation from the symptomatic leaves and shoots. Two white and mucoid colonies were isolated into the pure culture. Two isolates were identified as Erwinia amylovora according to the colony-polymerase chain reaction (PCR) with amsB primers and the phylogenetic tree using 16S rRNA sequences. To test of pathogenicity of two isolates, we inoculated immature pear fruits and understock of apple. We observed necrosis and oozes on immature pear fruits and shoot blight resulting in necrosis on apple shoots six days after inoculation. Colonies were recovered from the inoculated pears and apples, and identity was confirmed through colony PCR for amsB genes. To our knowledge, E. amylovora was first reported on Korean mountain ash native to South Korea.

Characterization of broad bean wilt virus 2 isolated from Perilla frutescens in Korea (국내 잎들깨에서 발생한 잠두위조바이러스2의 특성 구명)

  • Hyun-Sun Kim;Hee-Seong Byun;You-Ji Choi;Hyun-Yong Choi;Jang-Kyun Seo;Hong-Soo Choi;Bong-Choon Lee;Mikyeong Kim;Hae-Ryun Kwak
    • Korean Journal of Environmental Biology
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    • v.41 no.1
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    • pp.1-13
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    • 2023
  • Broad bean wilt virus 2 (BBWV2) is a species in the genus Fabavirus and family Secoviridae, which is transmitted by aphids and has a wide host range. The BBWV2 genome is composed of two single-stranded, positive-sense RNAs, RNA-1 and RNA-2. The representative symptoms of BBWV2 are mosaic, mottle, vein clearing, wilt, and stunting on leaves, and these symptoms cause economic damage to various crops. In 2019, Perilla fructescens leaves with mosaic and yellowing symptoms were found in Geumsan, South Korea. Reverse-transcription polymerase chain reaction (RT-PCR) was performed with specific primers for 10 reported viruses, including BBWV2, to identify the causal virus, and the results were positive for BBWV2. To characterize a BBWV2 isolate (BBWV2-GS-PF) from symptomatic P. fructescens, genetic analysis and pathogenicity tests were performed. The complete genomic sequences of RNA-1 and RNA-2 of BBWV2-GS-PF were phylogenetically distant to the previously reported BBWV2 isolates, with relatively low nucleotide sequence similarities of 76-80%. In the pathogenicity test, unlike most BBWV2 isolates with mild mosaic or mosaic symptoms in peppers, the BBWV2-GS-PF isolate showed typical ring spot symptoms. Considering these results, the BBWV2-GS-PF isolate from P. fructescens could be classified as a new strain of BBWV2.

High-efficiency deep geological repository system for spent nuclear fuel in Korea with optimized decay heat in a disposal canister and increased thermal limit of bentonite

  • Jongyoul Lee;Kwangil Kim;Inyoung Kim;Heejae Ju;Jongtae Jeong;Changsoo Lee;Jung-Woo Kim;Dongkeun Cho
    • Nuclear Engineering and Technology
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    • v.55 no.4
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    • pp.1540-1554
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    • 2023
  • To use nuclear energy sustainably, spent nuclear fuel, classified as high-level radioactive waste and inevitably discharged after electricity generation by nuclear power plants, must be managed safely and isolated from the human environment. In Korea, the land area is limited and the amount of high-level radioactive waste, including spent nuclear fuels to be disposed, is relatively large. Thus, it is particularly necessary to maximize disposal efficiency. In this study, a high-efficiency deep geological repository concept was developed to enhance disposal efficiency. To this end, design strategies and requirements for a high-efficiency deep geological repository system were established, and engineered barrier modules with a disposal canister for pressurized water reactor (PWR)-type and pressurized heavy water reactor type Canada deuterium uranium (CANDU) plants were developed. Thermal and structural stability assessments were conducted for the repository system; it was confirmed that the system was suitable for the established strategies and requirements. In addition, the results of the nuclear safety assessment showed that the radiological safety of the new system met the Korean safety standards for disposal of high-level radioactive waste in terms of radiological dose. To evaluate disposal efficiency in terms of the disposal area, the layout of the developed disposal areas was assessed in terms of thermal limits. The estimated disposal areas were 2.51 km2 and 1.82 km2 (existing repository system: 4.57 km2) and the excavated host rock volumes were 2.7 Mm3 and 2.0 Mm3 (existing repository system: 4.5 Mm3) for thermal limits of 100 ℃ and 130 ℃, respectively. These results indicated that the area and the excavated volume of the new repository system were reduced by 40-60% compared to the existing repository system. In addition, methods to further improve the efficiency were derived for the disposal area for deep geological disposal of spent nuclear fuel. The results of this study are expected to be useful in establishing a national high-level radioactive waste management policy, and for the design of a commercial deep geological repository system for spent nuclear fuels.

An Importance-Performance Analysis of Location Selection Factors for International Distribution Center in Port Hinterland (IPA기법을 통한 항만배후단지 내 국제물류센터 입주결정요인 분석)

  • Kim, Si-Hyun
    • Korea Trade Review
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    • v.42 no.1
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    • pp.283-301
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    • 2017
  • As a consequence of the changed role and functions in port operations, the role of port hinterland has transformed to multi-functional logistic centre linking more efficiently elements of the supply chain. This paper analysed distribution centre selection factors in Busan new port hinterland, aiming to diagnose and evaluate the operational situations of port hinterland as multi-functional logistics centre. Based on a data collected from all 122 samples located in Busan new port hinterland, determinants for location competitiveness identified were: political support, market potentiality, infrastructure utilization, market niche, and connectivity. Comparing the difference between an importance and performance, it is revealed that the target port hinterland requires urgent improvement in political supports such as incentive programmes offered by host country, free trade system and related law, financial assistance in constructing distribution centers, and simplicity, ease and efficiency of administrative procedures. The results provide useful insights for establishing future improvement strategies and a strategic agenda to successfully respond to the demands of the companies located in port hinterlands and/or new customers those who want to move in.

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Biological Control of Stem Rot of Pepper caused by Sclerotium rolfsii using by Bacillus amyloliquefaciens KBC1009 (길항세균 Bacillus amyloliquefaciens KBC1009를 이용한 고추 흰비단병의 생물학적 방제)

  • Kang, Jae-Gon;Lee, Young-Ui;Park, Jeong-chan;Jeong, Yoon-Woo;Park, Chang-Seuk;Kang, Hoon-Serg
    • Journal of agriculture & life science
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    • v.50 no.4
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    • pp.27-34
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    • 2016
  • Sclerotium rolfsii is a well known broad host range soil borne plant pathogenic fungus and caused serious damage to various vegetable crops. To develop an effective biological control agent for S. rolfsii, an isolate which showed strong inhibitory effect on the mycelial growth of S. rolfsii was selected among the antagonistic bacterial isolates collected from vinyl-house soil. The bacterial isolate was identified as Bacillus amyloliquefaciens KBC1009 based on the morphological, physiological characteristics and by 16S rRNA sequence analysis. The growth conditions for B. amyloliquefaciens KBC1009 were optimized in LB media(pH7) by culturing at 30℃ for 72 hrs. Glucose and yeast extract were confirmed as the best carbon and nitrogen sources, respectively. In order to test the inhibitory effect of B. amyloliquefaciens KBC1009 to stem rot of pepper, green house experiment was conducted. Drench of 1/500 diluted bacterial suspension of B. amyloliquefaciens KBC1009(5×108 cfu/ml) to each pepper plant 3 times with 10 days interval showed 66.7% control effectiveness. These results suggest that B. amyloliquefaciens KBC1009 is one of promising biocontrol agent to control stem rot caused by Sclerotium rolfsii.

Study on the Non-Characteristic Space Concept of Korean Traditional Residential Space Shown in Public Space of Sharehouse (셰어하우스 공용 공간에 나타난 한국전통주거공간의 무자성적(無自性的) 특성에 관한 연구 - 일본 셰어하우스 공용공간의 사례분석을 중심으로 -)

  • Yoon, Deuk Geun;Kim, Kai Chun
    • Korea Science and Art Forum
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    • v.19
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    • pp.515-525
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    • 2015
  • Along with a rapid increase in the number of 1-person households where the concept of 1-person consumption and 1-person economy is emphasized, the proportion of 1-person household in Korea occupies a 25.3% at present, and the number is expected to grow continuously. And this rapid increase will bring about a host of problems such as housing shortage in the metropolitan area, egoism, loneliness felt by individuals, and other social problems such as crime. As an alternate movement to this phenomenon, concern on and interest in community, community culture, and sharing of space are on the rise, together with concern on and preference for sharehouse culture. In news media as well, articles on life shared with members of a sharehouse often appear. This sharehouse, which is widely spread and well received in Japan, not only reduces economic burden but also creates their own community and promotes their own culture. In this sense, it is a new way of life that represents benefits of sharing, well beyond just economic interests. Accordingly, In this research, an attempt was made tp examine the circumstances in the use of space based on existing studies on sharehouse characteristics in order to shed new lights on the meaning that non-characteristic space concept of the traditional residential space has as the concept of sharehouse space by considering it in connection with the non-characteristic space concept of the Korean traditional residential space where diverse circumstances occur centered on communal life and which were accepted by all naturally.