• Title/Summary/Keyword: 차세대 서열 분석

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Genomic epidemiology and surveillance of zoonotic viruses using targeted next-generation sequencing (표적화 차세대염기서열분석법을 이용한 인수공통 바이러스의 유전체 역학과 예찰)

  • Seonghyeon Lee;Seung-Hwan Baek;Shivani Rajoriya;Sara Puspareni;Won-Keun Kim
    • Korean Journal of Veterinary Service
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    • v.46 no.1
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    • pp.93-106
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    • 2023
  • Emerging and re-emerging zoonotic viruses become critical public health, economic, societal, and cultural burdens. The Coronavirus disease-19 (COVID-19) pandemic reveals needs for effective preparedness and responsiveness against the emergence of variants and the next virus outbreak. The targeted next-generation sequencing (NGS) significantly contributes to the acquisition of viral genome sequences directly from clinical specimens. Using this advanced NGS technology, the genomic epidemiology and surveillance play a critical role in identifying of infectious source and origin, tracking of transmission chains and virus evolution, and characterizing the virulence and developing of vaccines during the outbreak. In this review, we highlight the platforms and preparation of targeted NGS for the viral genomics. We also demonstrate the application of this strategy to take advantage of the responsiveness and prevention of emerging zoonotic viruses. This article provides broad and deep insights into the preparedness and responsiveness for the next zoonotic virus outbreak.

Correlation between Disease Occurrences and Microbial Community Structure by Application of Organic Materials in Pepper (유기농자재 사용에 따른 고추 병해 발생과 토양 미생물상 구조의 상관관계)

  • Cho, Gyeongjun;Kim, Seong-Hyeon;Lee, Yong-Bok;Kwak, Youn-Sig
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.202-209
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    • 2020
  • Organic farming is necessary to sustainable agriculture, preserve biodiversity and continued growth the sector in agriculture. In organic farming, reduced usage of chemical agents that adversely affect human health and environment, employing amino acids and oil cake fertilizer, plant extracts, and microbial agents are used to provide safe agricultural products to consumers. To investigation microbiome structure, we proceeded on the pepper plant with difference fertilizers and treatments in organic agriculture for three years. The microbial communities were analyzed by the next generation sequencing approach. Difference soil microbiota communities were discovered base on organic fertilizer agents. Occurrences of virus and anthracnose diseases had a low incidence in conventional farming, whereas bacteria wilt disease had a low incidence in microbial agents treated plots. Microbe agents, which applied in soil, were detected in the microbial community and the funding suggested the applied microbes successfully colonized in the organic farming environment.

Identification of Uncharacterized Anti-microbial Peptides Derived from the European Honeybee (꿀벌 Apis mellifera에서 유래 한 특성화 되지 않은 항균성 펩티드의 동정)

  • Park, Hee Geun;Kim, Dong Won;Lee, Man-Young;Choi, Yong Soo
    • Journal of Life Science
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    • v.30 no.1
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    • pp.64-69
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    • 2020
  • The European honeybee (Apis mellifera L.) has multiple anti-microbial peptides, but many were unknown and demands for their characterization have increased. This study therefore focused on identifying novel anti-microbial peptides (AMPs) from A. mellifera L. To obtain high-throughput transcriptome data of the honeybee, we implemented next-generation sequencing (NGS), isolating novel AMPs from total RNA, and generated 15,314 peptide sequences, including 44 known, using Illumina HiSeq 2500 technology. The uncharacterized peptides were identified based on specific features of possible AMPs predicted in the sequencing analysis. AMP5, one such uncharacterized peptide, was expressed in the epidermis, body fat, and venom gland of the honeybee. We chemically synthesized this peptide and tested its anti-bacterial activity against Gram-negative Escherichia coli (KACC 10005) and Gram-positive Bacillus thuringiensis (KACC 10168) by anti-microbial assay. AMP5 exhibited anti-bacterial activity against E. coli (MIC50=22.04±0.66 μM) but not against B. thuringiensis. When worker bees were injected with E. coli, AMP5 was up-regulated in the body fat. This study therefore identified AMP5 in adult European honeybees and confirmed its anti-bacterial activity against Gram-negative E. coli.

Microbial diversity and physicochemical properties of takju and yakju (탁주와 약주의 이화학적 특성 및 미생물 군집 분석)

  • Koo, Ok Kyung;Lim, Eun Seob;Lee, Ae-Ran;Kim, Tae Wan
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.400-406
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    • 2018
  • Takju and yakju are traditional Korean alcoholic beverages that are prepared by fermentation of glutinous rice with nuruk, a cereal starter containing various bacteria, fungi, and yeast. In this study, physicochemical and microbial properties of a total of 12 commercial takju and yakju samples were analyzed; their pH, sweetness, and alcohol content were varied, depending on the type of alcohol, from pH 3.64-4.8, $5.1-24.8^{\circ}Bx$, and 4.6-18.5%, respectively. Microbial communities were analyzed with 16S rRNA amplicon sequencing using MiSeq system. At the phylum level, Firmicutes (86.2%) was the most dominant, followed by Proteobacteria (8.08%), Actinobacteria (2.56%), and Cyanobacteria (3.13%). Lactic acid bacteria, including Lactobacillus, Lactococcus, Leuconostoc, and Weissella were also frequently detected. Among eukaryotes, Saccharomyces cerevisiae was the most dominant in these samples.

Phylogenetic characteristics of actinobacterial population in bamboo (Sasa borealis) soil (조릿대 대나무림 토양 내 방선균군집의 계통학적 특성)

  • Lee, Hyo-Jin;Han, Song-Ih;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.59-64
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    • 2016
  • In this study, a pyrosequencing was performed and analyzed to verify the phylogenetic diversity of actinomycetes in the bamboo (Sasa borealis) soil as a base study to obtain the genetic resources of actinomycetes. It was found that the rhizosphere soil had much various distribution in bacterial communities showing a diversity of 8.15 with 2,868 OTUs, while the litter layer showed a diversity of 7.55 with 2,588 OTUs. The bacterial community in the bamboo soil was composed of 35 phyla and the predominant phyla were Proteobacteria (51-60%), Bacteroidetes (16-20%), Acidobacteria (4-16%) and Actinobacteria (4-14%). In particular, Actinobacteria including Micromonosporaceae and Streptomycetaceae had a diverse distribution of actinomycetes within the six orders, 35 families and 121 genera, and it was characterized that about 83% of actinomycetes within Actinomycetales belonged to the 28 families. Among the dominant actinobacterial populations, Micromonosporaceae, Pseudonocardiaceae and Streptomycetaceae were representative family groups in the bamboo soils.

Analysis of Gene Expression in Larval Fat Body of Plutella Xylostella Under High Temperature (고온에서 배추좀나방 유충 지방체의 유전자 발현 변화 분석)

  • Kim, Kwang Ho;Lee, Dae-Weon
    • Korean Journal of Environmental Agriculture
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    • v.37 no.4
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    • pp.324-332
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    • 2018
  • BACKGROUND: Insects are ectothermic organisms in terrestrial ecosystems and play various roles such as controlling plant biomass and maintaining species diversity. Because insects are ectothermic, their physiological responses are very sensitive to environmental temperature which determines survival and distribution of insect population and that affects climate change. This study aimed to identification of genes contributing to fitness under high temperature. METHODS AND RESULTS: To identify genes contributing to fitness under high temperature, the transcriptomes of fat body in Plutella xyostella larva have been analyzed via next generation sequencing. From the fat body transcriptomes, structure-related proteins, heat shock proteins, antioxidant enzymes and detoxification proteins were identified. Genes encoding proteins such as structural proteins (cuticular proteins, chitin synthase and actin), stress-related protein (cytochrome P450), heat shock protein and antioxidant enzyme (catalase) were up-regulated at high temperature. In contrast expression of glutathione S transferase was down-regulated. CONCLUSION: Identifications of temperature-specific up- or down-regulated genes can be useful for detecting temperature adaptation and understanding physiological responses in insect pests.

A comparison of the reproduction of two closely related species, tiger worm(Eisenia fetida) and red tiger worm(Eisenia andrei) when the organic sludge was suppied to them (유기성 슬러지 먹이에 대한 두 근연종인 줄지렁이(Eisenia fetida)와 붉은줄지렁이(Eisenia andrei)의 생식반응 비교)

  • Bae, Yoon-Hwan;Shin, Hyun-Gon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.29 no.3
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    • pp.27-33
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    • 2021
  • CO I gene sequence analysis was applied to earthworms that had been used as test animals in toxicity test in Institute of Kyeongbook Agrochemicals and earthworms used as vermicomposting agents in the farm of Youngdong province to identify their species names. In terms of molecular species, the former was identified as Eisenia fetida and the latter was Eisenia andrei. Cocoons produced from Eisenia fetida was more than those from Eisenia andrei. And No. of adults developed from eggs of Eisenia fetida was more or less higher than those developed from eggs of Eisenia andrei. These results were contradictory to previous reports on two Eisenia spp.. When Eisenia fetida was crossed with Eisenia andrei, hybridized eggs were produced and adults were developed from those eggs, but cocoons and adults were much less than those from non-crossed Eisenia fetida or Eisenia andrei. This indicated that two Eisenia spp. were not distinctly different biological species because there was no complete 'reproductive isolation' between Eisenia fetida and Eisenia andrei. However, this also meant that Eisenia fetida and Eisenia andrei had already been on the tract of speciation.

Development of Chloroplast Genome-based Insertion/Deletion Markers in the Genus Broussonetia (닥나무 속 식물의 엽록체 유전체 기반 InDel 마커의 개발)

  • Eun Jee Lee;Yoon A Kim;Mi Sun Lee;Ju Hyeok Kim;Young Kyu Choi;Jung Sung Kim;Chang Seob Sin;Yi Lee
    • Korean Journal of Plant Resources
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    • v.36 no.4
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    • pp.290-298
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    • 2023
  • Several members of the genus Broussonetia are woody plants with high-quality cellulose fibers and are used to make a traditional type of Korean paper known as Hanji. Three of these species, Broussonetia kazinoki, Broussonetia monoica, and Broussonetia papyrifera, are found in the Korean Peninsula. Because it is challenging to distinguish different Broussonetia species based on morphology alone, we have developed a set of insertion/deletion (InDel) markers for genetic identification of these species. From twenty-two Broussonetia samples collected throughout Korea, we selected six for next-generation sequencing analysis. InDel marker candidates were identified by comparing this sequence information with the B. kazinoki chloroplast genome sequence. The marker candidates were used to screen the genomes of the twenty-two Broussonetia plants, and five useful chloroplast-based InDel markers were identified. Detailed genotyping using these five markers showed that the twenty-two plants of the genus Broussonetia could be clustered into five groups, verifying that the markers developed here can be used for breeding, identification, and analysis of species in the genus Broussonetia.

A Bioconversion Study on the Zanthoxylum schinfolium by Fermenting Bacteria and Their Functional Enhancement (유용 발효미생물 활용 생물전환 공정을 활용한 산초열매의 기능성 증대 방안 연구)

  • Lim, Jeong-Muk;Lee, Se-Won;Lee, Seong-Hyeon;Lee, Jeong-Ho;Oh, Byung-Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.64-64
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    • 2018
  • 생물전환(Bioconversion)은 천연소재의 기능성을 증대시키기 위한 방안으로 많은 연구가 진행되고 있으며 다양한 산업에서도 활용되고 있어 유용미생물을 기반 차세대 기술로 각광받고 있다. 이러한 기술의 도입은 식품은 물론 의약품 및 화장품 산업에서도 활발히 사용되고 있으며, 특히 최근 기능성 제품에 대한 소비가 급증함에 따라 그 중요성이 높아지고 있다. 산초(Zanthoxylum schinfolium)는 limonene, citronellal, phellandrene 등의 다양한 유효물질을 함유하고 있으며 유효성분들로부터 유래되는 항산화 활성과 항암활성, 항균활성 등의 효능을 지니는 것으로 보고된바 있다. 본 연구의 생물전환 공정에 사용된 유산균들은 전통 발효식품으로 알려진 다양한 젓갈류로부터 분리하였으며, 16S rDNA 염기서열 분석을 통해 유전학적 특성을 확인하였다. 또한 확보된 유산균들을 사용하여 산초(전북 진안군) 분말의 발효공정을 수행하였으며, 산초의 최적 추출조건을 선정하고 추출물을 제조하여 생물전환 공정 전 후 활성의 변화추이를 관찰하였다. 추출물의 활성평가는 항산화 효능 및 유효성분 함량을 평가하기 위하여 DPPH radical scavenging activity와 total polyphenol 함량을 평가하고 세포주를 활용해 MTT assay, Nitric oxide(NO) 생성억제 효능을 확인하여 세포독성 및 항염증 활성을 확인하였다. 실험결과, 생물전환 공정에 사용할 유용 미생물을 확보하기 위한 실험을 통해 다양한 젓갈류에서 다양한 미생물을 확보할 수 있었으며 약 16종의 유산균을 분리하였다. 분리된 미생물을 사용하여 산초 분말의 생물전환 공정을 실시한 결과, 5종의 유용미생물 처리에서 무처리 대비 DPPH radical 소거능 및 polyphenol 함량이 유의적으로 증가됨을 확인할 수 있었다. 그 중 가장 높은 활성을 나타내는 균주를 16S rDNA 염기서열 분석을 통해 확인한 결과 Weissella confusa D1로 확인되었다. 선별 균주를 활용한 생물전환 공정 후 항산화 활성은 대조군 대비 약 120%의 활성을 나타냈으며, polyphenol의 함량은 약 126%로 증가하는 것을 확인할 수 있었다. 더 나아가 생물전환 공정 후 산초추출물의 세포독성은 처리전과 비교하여 월등히 감소하는 경향을 확인할 수 있었으며, 항염효능 또한 증가하는 경향을 나타내는 것이 확인되었다.

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Phylogenetic characteristics of bacterial populations and isolation of aromatic compounds utilizing bacteria from humus layer of oak forest (상수리림 부식층으로부터 방향족 화합물 분해세균의 분리 및 세균군집의 계통학적 특성)

  • Han, Song-Ih
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.175-182
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    • 2016
  • In this study, we isolated aromatic compounds (lignin polymers) utilizing bacteria in humus layer of oak forest and investigated phylogenetic characteristics and correlation with major bacterial populations in the humus layer by pyrosequencing. Forty-two isolates using aromatic compounds such as p-anisic acid, benzoic acid, ferulic acid and p-coumaric acid were isolated and phylogentic analyses based on 16S rRNA gene sequences showed that the isolates belonged to the genus Rhizobium, Sphingomonas, Burkhorlderia, and Pseudomonas. Among these, Burkhorlderia species which belong to Betaproteobacteria class occupied 83% among the isolates. The bacterial populations in humus layer of oak forest were characterized by next generation pyrosequencing based on 16S rRNA gene sequences. The humus sample produced 7,862 reads, 1,821 OTUs and 6.76 variability index with 97% of significance level, respectively. Bacterial populations consist of 22 phyla and Betaproteobacteria were the major phylum consisting of 15 genera including Burkholderia, Polaromonas, Ralstoria, Zoogloea, and Variovorax. Approximately fifty percentage of them was Burkholderia. Burkholderia as the majority of population in the humus was considered to play a role in degrading lignin in humus layer of oak forest.