• 제목/요약/키워드: Sequencing problem

검색결과 218건 처리시간 0.031초

Molecular Analysis of Carbapenem-Resistant Enterobacteriaceae at a South Korean Hospital

  • Lee, Miyoung;Choi, Tae-Jin
    • 한국미생물·생명공학회지
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    • 제48권3호
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    • pp.389-398
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    • 2020
  • The prevalence of carbapenem-resistant Enterobacteriaceae (CRE) is increasing globally, resulting in high mortality rates. Although CRE is a relatively recent problem in Korea (the first case was not diagnosed until 2010), it is responsible for serious morbidities at an alarming rate. In this study, we carried out a molecular genetic analysis to determine the incidence of CRE and carbapenemase-producing Enterobacteriaceae (CPE) at a general hospital in Korea between August 2017 and August 2019. Forty strains of CPE were isolated from various clinical specimens and analyzed via antimicrobial susceptibility testing, polymerase chain reaction to detect β-lactamase genes, deoxyribonucleic acid sequencing, multilocus sequence typing, curing testing, and conjugal transfer of plasmids. The results demonstrated that all 40 isolates were multidrug-resistant. The fluoroquinolone susceptibility test showed that 75% of the Enterobacteriaceae isolates were resistant to ciprofloxacin, whereas 72.5% were resistant to trimethoprim-sulfamethoxazole. Further, conjugation accounted for 57.5% of all resistant plasmid transfer events, which is 4.3-fold higher than that observed in 2010 by Frost et al. Finally, the high detection rate of transposon Tn4401 was associated with the rapid diffusion and evolution of CPE. Our results highlight the rapid emergence of extensively drugresistant strains in Korea and emphasize the need for employing urgent control measures and protocols at the national level.

튜브 제조 시스템의 생산 스케줄링 사례연구 (A Case Study on the Scheduling for a Tube Manufacturing System)

  • 임동순;박찬현;조남찬;오현승
    • 산업경영시스템학회지
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    • 제32권3호
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    • pp.110-117
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    • 2009
  • This paper introduces a case study for efficient generation of production schedules in a tube manufacturing system. The considered scheduling problem consists of two sub problems : lot sizing for a job and Job sequencing. Since these problems require simulation optimization in which the performance measures are obtained by simulation execution, the trade-off between solution quality and computation time is an important issue. In this study, the optimal lot size for every product type is determined from simulation experiments. Then, target production quantity for each product type is transformed to several jobs such that a Job consists of determined lot size. To obtain the good solution for a Job sequence in a reasonable time, a number of alternatives are generated from heuristic rules developed by intuition and analysis of the considered system, and a job sequence is selected from simulation experiments.

Method of DNA Extraction from Pinus rigida Wood Pretreated with Sandpaper

  • Lee, Jamin;Kim, Tae-Jong
    • Journal of the Korean Wood Science and Technology
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    • 제46권4호
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    • pp.402-414
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    • 2018
  • Species identification of wood provides important information for archaeology, restoration of cultural assets, preventing illegal logging, and more. Wood species are usually identified based on their anatomical features with the use of a microscope. However, this method may not be able to distinguish between anatomically similar species or subspecies. To overcome this problem, wood species need to be identified at the molecular level using DNA sequencing. However, unlike living plant cells, wood is difficult to pulverize using a mortar, and DNA extraction from dried wood is challenging. To solve these problems, we propose a pretreatment method in which wood is pulverized using 60-grit sandpaper and hydrated with water for 2 days. Using this method, we were able to stably amplify the rpoB gene from the extracted DNA of Pinus rigida. In addition, sequence analysis of the rpoB gene revealed six single nucleotide polymorphisms (SNPs), which classified the rpoB sequences in the genus Pinus into five groups. Our data indicate that although these SNPs were not suitable for species identification, they can potentially be used to determine the origin of different wood subspecies or individual samples of wood.

A Rapid and Simple Method for Construction and Expression of a Synthetic Human Growth Hormone Gene in Escherichia coli

  • Roytrakul, Sittiruk;Eurwilaichitr, Lily;Suprasongsin, Chittiwat;Panyim, Sakol
    • BMB Reports
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    • 제34권6호
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    • pp.502-508
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    • 2001
  • A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

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Computational Detection of Prokaryotic Core Promoters in Genomic Sequences

  • Kim Ki-Bong;Sim Jeong Seop
    • Journal of Microbiology
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    • 제43권5호
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    • pp.411-416
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    • 2005
  • The high-throughput sequencing of microbial genomes has resulted in the relatively rapid accumulation of an enormous amount of genomic sequence data. In this context, the problem posed by the detection of promoters in genomic DNA sequences via computational methods has attracted considerable research attention in recent years. This paper addresses the development of a predictive model, known as the dependence decomposition weight matrix model (DDWMM), which was designed to detect the core promoter region, including the -10 region and the transcription start sites (TSSs), in prokaryotic genomic DNA sequences. This is an issue of some importance with regard to genome annotation efforts. Our predictive model captures the most significant dependencies between positions (allowing for non­adjacent as well as adjacent dependencies) via the maximal dependence decomposition (MDD) procedure, which iteratively decomposes data sets into subsets, based on the significant dependence between positions in the promoter region to be modeled. Such dependencies may be intimately related to biological and structural concerns, since promoter elements are present in a variety of combinations, which are separated by various distances. In this respect, the DDWMM may prove to be appropriate with regard to the detection of core promoter regions and TSSs in long microbial genomic contigs. In order to demonstrate the effectiveness of our predictive model, we applied 10-fold cross-validation experiments on the 607 experimentally-verified promoter sequences, which evidenced good performance in terms of sensitivity.

전달손실 최대화를 위한 다층 흡음재-패널 배열 최적설계 (Optimization of Multilayered Foam-panel Sequence for Sound Transmission Loss Maximization)

  • 김용진;이중석;강연준;김윤영
    • 한국소음진동공학회논문집
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    • 제18권12호
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    • pp.1262-1269
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    • 2008
  • Though multilayered foam-panel structures have been widely used to reduce sound transmission in various fields, most of the previous works to design them were conducted by repeated analyses or experiments based on initially given configurations or sequences. Therefore, it was difficult to obtain an optimal sequence of multilayered foam-panel structure yielding superior sound isolation capability. In this work, we propose a new design method to sequence a multi-panel structure lined with a poroelastic material having maximized sound transmission loss. Being formulated as a one-dimensional topology optimization problem fur a given target frequency, the optimal sequencing of panel-poroelastic layers is systematically carried out in an iterative manner. In this method, a panel layer is expressed as a limiting case of a poroelastic layer to facilitate the optimization process. This means that main material properties of a poroelastic material are treated as interpolated functions of design variable. The designed sequences of panel-poroelastic multilayer were shown to be significantly affected by the target frequencies; more panels were obtained at higher target frequency. The sound transmission loss of the system was calculated by the transfer matrix derived from Biot's theory.

Identification of Vibrio species isolated from cultured olive flounder (Paralichthys olivaceus) in Jeju Island, South Korea

  • Sohn, Hanchang;Kim, Jeongeun;Jin, Changnam;Lee, Jehee
    • Fisheries and Aquatic Sciences
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    • 제22권7호
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    • pp.14.1-14.8
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    • 2019
  • Olive flounder (Paralichthys olivaceus) is the major species developed for aquaculture in South Korea. Over the long history of olive flounder aquaculture, complex and diverse diseases have been a major problem, negatively impacting industrial production. Vibriosis is a prolific disease which continuously damages olive flounder aquaculture. A bacterial disease survey was performed from January to June 2017 on 20 olive flounder farms on Jeju Island. A total of 1710 fish were sampled, and bacteria from the external and internal organs of 560 fish were collected. Bacterial strains were identified using 16 s rRNA sequencing. Twenty-seven species and 184 strains of Vibrio were isolated during this survey, and phylogenetic analysis was performed. Bacterial isolates were investigated for the distribution of pathogenic and non-pathogenic species, as well as bacterial presence in tested organs was characterized. V. gigantis and V. scophthalmi were the dominant non-pathogenic and pathogenic strains isolated during this survey, respectively. This study provides data on specific Vibrio spp. isolated from cultured olive flounder in an effort to provide direction for future research and inform aquaculture management practices.

Influence of Companion Planting on Microbial Compositions and Their Symbiotic Network in Pepper Continuous Cropping Soil

  • Jingxia Gao;Fengbao Zhang
    • Journal of Microbiology and Biotechnology
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    • 제33권6호
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    • pp.760-770
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    • 2023
  • Continuous cropping obstacles have become a serious factor restricting sustainable development in modern agriculture, while companion planting is one of the most common and effective methods for solving this problem. Here, we monitored the effects of companion planting on soil fertility and the microbial community distribution pattern in pepper monoculture and companion plantings. Soil microbial communities were analyzed using high-throughput sequencing technology. Companion plants included garlic (T1), oat (T2), cabbage (T3), celery (T4), and white clover (T5). The results showed that compared with the monoculture system, companion planting significantly increased the activities of soil urease (except for T5) and sucrase, but decreased catalase activity. In addition, T2 significantly improved microbial diversity (Shannon index) while T1 resulted in a decrease of bacterial OTUs and an increase of fungal OTUs. Companion planting also significantly changed soil microbial community structures and compositions. Correlation analysis showed that soil enzyme activities were closely correlated with bacterial and fungal community structures. Moreover, the companion system weakened the complexity of microbial networks. These findings indicated that companion plants can provide nutrition to microbes and weaken the competition among them, which offers a theoretical basis and data for further research into methods for reducing continuous cropping obstacles in agriculture.

넙치(Paralichthys olivaceus)자치어 장관백탁증(Bacterial white enteritis) 원인균의 신속 검출 (Rapid Detection of the pathogenic agent of Bacterial white enteritis of Larval and Juvenile Stages in Olive flounder (Paralichthys olivaceus))

  • 문영건;박근태;손홍주;이상현;이정민;허문수
    • 한국어병학회지
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    • 제17권3호
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    • pp.159-169
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    • 2004
  • 2003년 5월과 2003년 10월동안에 제주도내 5개소의 넙치 종묘배양장에서 초기 먹이로 공급 되어지는 동물성 플랑크톤인 rotifer와 20-30일령 넙치 자어에서 장관백탁증 원인균으로 알려진 V. ichthyoenteri를 분리하기 위해 실험한 결과 총 71개의 Vibrio sp. 분리가 되었고, 생화학적 동정결과 2개의 그룹에서 24개의 V ichthyoenteri가 동정 되었다. V. ichthyoenteri의 신속한 검출을 위한 종특이적 primer를 V. ichthyoenteri(KCCM 40870)ISR의 특이적인 서열을 이용하여 제작하였다. V. ichthyoenteri를 포함한 20종의 Vibrio속 균주의 genomic DNA와 18group 분리균주 genomic DNA를 PCR한 결과 V. ichthyoenteri 만의 특이적인 band가 생성됨을 알 수가 있다. 따라서 V. ichthyoenteri(KCCM 40870) ISR의 서열로 제작한 primer가 넙치 자치어에 발병하는 장관백탁증 원인균인 Vibrio ichthyoenteri의 신속한 검출과 정확한 동정을 할 수 있는 molecular marker로 이용할 수 있음을 확인하였다.

도시 내 육상 생물종 모니터링을 위한 환경DNA 리뷰 및 적용 (Review and application of environmental DNA (eDNA) investigation of terrestrial species in urban ecosystem)

  • 김휘문;김성열;박일수;이현정;김경태;김영;김혜정;곽민호;임태양;박찬;송원경
    • 한국환경복원기술학회지
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    • 제23권2호
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    • pp.69-89
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    • 2020
  • Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.