• 제목/요약/키워드: Sequencing Strategy

검색결과 93건 처리시간 0.025초

High-throughput Gene Expression Analysis to Investigate Host-pathogen Interaction in Avian Coccidiosis

  • Lillehoj Hyun, S.
    • 한국가금학회지
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    • 제34권1호
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    • pp.77-83
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    • 2007
  • Poultry products including meat and eggs constitute a major protein source in the American diet and disease-causing pathogens represent major challenges to the poultry industry. More than 95% of pathogens enter the host through the mucosal surfaces of the respiratory, digestive and reproductive tracts and over the past few decades, the two main mechanisms used to control diseases have been the use of vaccines and antibiotics. However, in the poultry industry, there are mounting concerns over the ability of current vaccines to adequately protect against emerging hyper-virulent strains of pathogens and a lack of suitable, cost effective adjuvants. Thorough investigation of the immunogenetic responses involved in host-pathogen interactions will lead to the development of new and effective strategies for improving poultry health, food safety and the economic viability of the US poultry industry. In this paper, I describe the development of immunogenomic and proteomic tools to fundamentally determine and characterize the immunological mechanisms of the avian host to economically significant mucosal pathogens such as Eimeria. Recent completion of poultry genome sequencing and the development of several tissue-specific cDNA libraries in chickens are facilitating the rapid application of functional immunogenomics in the poultry disease research. Furthermore, research involving functional genomics, immunology and bioinformatics is providing novel insights into the processes of disease and immunity to microbial pathogens at mucosal surfaces. In this presentation, a new strategy of global gene expression using avian macrophage (AMM) to characterize the multiple pathways related to the variable immune responses of the host to Eimeria is described. This functional immunogenomics approach will increase current understanding of how mucosal immunity to infectious agents operates, and how it may be enhanced to enable the rational development of new and effective strategies against coccidiosis and other mucosal pathogens.

Artificial induction and isolation of cadmium-tolerant soil bacteria

  • Lee, Sangman
    • Journal of Applied Biological Chemistry
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    • 제63권2호
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    • pp.125-129
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    • 2020
  • Environmental pollution caused by various heavy metals is a serious global problem. To solve this problem, microbial bioremediation of contaminated metals has developed rapidly as an effective strategy when physical and chemical techniques are not suitable. In this study, cadmium (Cd)-tolerant soil bacteria were isolated via artificial induction in laboratory conditions instead of screening bacteria naturally adapted to metal-contaminated soils. Wild-type (WT) bacteria grown in uncontaminated soils were artificially and sequentially adapted to gradually increasing Cd concentrations of up to 15 mM. The resultant cells, named Soil-CdR15, survived at a Cd concentration of 10 mM, whereas WT cells failed to survive with 4 mM Cd on solid media for 2 d. In liquid media containing Cd, the SoilCdR15 cells grew with 15 mM Cd for 7 d, whereas the WT cells could not grow with 5 mM Cd. Both Soil-CdR15 and WT cells removed approximately 35% of Cd at the same capacity from liquid media containing either 0.5 or 1.0 mM Cd over 2 d. In addition to Cd, the Soil-CdR15 cells showed increased resistance to nickel, zinc, and arsenic compared to WT cells. The Soil-CdR cells were identified as Burkholderia sp. by partial sequencing of 16S rRNA. The data presented in this study demonstrate that isolation of heavy metal-tolerant microorganisms via artificial induction in laboratory conditions is possible and may be useful for the application of the microorganisms for the bioremediation of heavy metals.

Sampling and Selection Factors that Enhance the Diversity of Microbial Collections: Application to Biopesticide Development

  • Park, Jun-Kyung;Lee, Seung-Hwan;Lee, Jang-Hoon;Han, Songhee;Kang, Hunseung;Kim, Jin-Cheol;Kim, Young Cheol;McSpadden Gardener, Brian
    • The Plant Pathology Journal
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    • 제29권2호
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    • pp.144-153
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    • 2013
  • Diverse bacteria are known to colonize plants. However, only a small fraction of that diversity has been evaluated for their biopesticide potential. To date, the criteria for sampling and selection in such bioprospecting endeavors have not been systematically evaluated in terms of the relative amount of diversity they provide for analysis. The present study aimed to enhance the success of bioprospecting efforts by increasing the diversity while removing the genotypic redundancy often present in large collections of bacteria. We developed a multivariate sampling and marker-based selection strategy that significantly increase the diversity of bacteria recovered from plants. In doing so, we quantified the effects of varying sampling intensity, media composition, incubation conditions, plant species, and soil source on the diversity of recovered isolates. Subsequent sequencing and high-throughput phenotypic analyses of a small fraction of the collected isolates revealed that this approach led to the recovery of over a dozen rare and, to date, poorly characterized genera of plant-associated bacteria with significant biopesticide activities. Overall, the sampling and selection approach described led to an approximately 5-fold improvement in efficiency and the recovery of several novel strains of bacteria with significant biopesticide potential.

자동화 컨테이너 터미널에서 이적작업을 위한 장치장 크레인 작업할당 (Yard Crane Dispatching for Remarshalling in an Automated Container Terminal)

  • 배종욱;박영만
    • 한국항해항만학회지
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    • 제36권8호
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    • pp.665-671
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    • 2012
  • 이적작업은 자동화 컨테이너 터미널에서 하역작업의 생산성을 높이는 운영 전략으로 연구되고 있다. 이는 장치장 블록 내 컨테이너들의 재배치 작업을 의미한다. 이적작업의 크레인 작업할당 문제는 작업가능시간에 대한 제약을 고려하여 이적작업의 효과를 최대화하기 위해 가능한 이적작업 중에서 효율적인 이적작업을 선정하고 이들의 작업순서를 정하는 것이다. 본 연구는 혼합정수계획법을 활용하여 장치장 크레인의 작업할당 문제에 대한 최적화 수리모형을 개발하고 이의 예를 제시하였다. 그러나 수리모형은 계산시간이 많이 소요되어 현장에서 사용되기 어렵다. 따라서 현실 적용을 위해 대표적인 5개의 작업할당 규칙을 비교 평가하였다. 수치실험에서 maximum weight ratio(MR) 규칙이 전반적으로 뛰어난 성능을 보였다.

빅데이터 및 고성능컴퓨팅 프레임워크를 활용한 유전체 데이터 전처리 과정의 병렬화 (Parallelization of Genome Sequence Data Pre-Processing on Big Data and HPC Framework)

  • 변은규;곽재혁;문지협
    • 정보처리학회논문지:컴퓨터 및 통신 시스템
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    • 제8권10호
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    • pp.231-238
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    • 2019
  • 차세대 염기 서열 분석법이 생성한 유전체 원시 데이터를 기존의 방식대로 하나의 서버에서 분석하기 위해서는 데이터 크기에 따라 수십 시간이 필요할 수 있다. 그러나 응급 환자의 진단처럼 수 시간 내에 결과를 알아야 하는 상황이 존재하기 때문에 단일 유전체 분석의 성능을 향상시킬 필요가 있다. 본 연구에서는 빅데이터 기술의 병렬화 기법과 고속의 네트워크로 연결되고 병렬파일시스템을 공유하는 고성능컴퓨팅 클러스터를 적극적으로 활용하여 분석 시간을 크게 단축시킬 수 있는 유전체 데이터 분석의 전처리 프로세스의 병렬화 방법을 제안한다. 분석 데이터의 신뢰성을 위해 기존의 검증된 분석 도구 및 알고리즘을 새로운 환경에 맞게 병렬화 하는 전략을 선택하였다. 프로세스의 병렬화, 데이터의 분배 및 병렬 병합 기법을 개발하였고 실험을 통해 성능 향상을 확인하였다.

Effect of JAK-STAT pathway in regulation of fatty liver hemorrhagic syndrome in chickens

  • Zhu, Yaling;Mao, Huirong;Peng, Gang;Zeng, Qingjie;Wei, Qing;Ruan, Jiming;Huang, Jianzhen
    • Animal Bioscience
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    • 제34권1호
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    • pp.143-153
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    • 2021
  • Objective: To explore the molecular mechanisms of fatty liver hemorrhagic syndrome (FLHS) in laying hens, an experiment was conducted to reveal the differences in histopathological observation and gene expression between FLHS group and normal group. Methods: We compared the histopathological difference using hematoxylin and eosin staining and proceeded with RNA sequencing of adipose tissue to search differentially expressed genes and enriched biological processes and pathways. Then we validated the mRNA expression levels by real-time polymerase chain reaction and quantified protein levels in the circulation by enzyme-linked immunosorbent assay. Results: We identified 100 differentially expressed transcripts corresponding to 66 genes (DEGs) were identified between FLHS-affected group and normal group. Seven DEGs were significantly enriched in the immune response process and lipid metabolic process, including phospholipase A2 group V, WAP kunitz and netrin domain containing 2, delta 4-desaturase sphingolipid 2, perilipin 3, interleukin-6 (IL-6), ciliary neurotrophic factor (CNTF), and suppressor of cytokine signaling 3 (SOCS3). And these genes could be the targets of immune response and be involved in metabolic homeostasis during the process of FLHS in laying hens. Based on functional categories of the DEGs, we further proposed a model to explain the etiology and pathogenesis of FLHS. IL-6 and SOCS3 mediate inflammatory responses and the satiety hormone of leptin, induce dysfunction of Jak-STAT signaling pathway, leading to insulin resistance and lipid metabolic disorders. Conversely, CNTF may reduce tissue destruction during inflammatory attacks and confer protection from inflammation-induced insulin resistance in FLHS chickens. Conclusion: These findings highlight the therapeutic implications of targeting the JAK-STAT pathway. Inhibition of IL6 and SOCS3 and facilitation of CNTF could serve as a favorable strategy to enhance insulin action and improve glucose homoeostasis, which are of importance for treating obesity-related disorders for chickens.

Pathological Impact on the Phyllosphere Microbiota of Artemisia argyi by Haze

  • Zhang, Yu-Zhu;Jiang, De-Yu;Zhang, Chi;Yang, Kun;Wang, Huai-Fu;Xia, Xiu-Wen;Ding, Wei-Jun
    • Journal of Microbiology and Biotechnology
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    • 제31권4호
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    • pp.510-519
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    • 2021
  • The pathological impact of haze upon the phyllosphere microbiota awaits investigation. A moderate degree of haze environment and a clean control were selected in Chengdu, China. Artemisia argyi, a ubiquitously distributed and extensively applied Chinese herb, was also chosen for experiment. Total genome DNA was extracted from leaf samples, and for metagenome sequencing, an Illumina HiSeq 2500 platform was applied. The results showed that the gene numbers of phyllosphere microbiota derived from haze leaves were lower than those of the clean control. The phyllosphere microbiota derived from both haze and clean groups shared the same top ten phyla; the abundances of Proteobacteria, Actinomycetes and Anorthococcuso of the haze group were substantially increased, while Ascomycetes and Basidiomycetes decreased. At the genus level, the abundances of Nocardia, Paracoccus, Marmoricola and Knoelia from haze leaves were markedly increased, while the yeasts were statistically decreased. KEGG retrieval demonstrated that the functional genes were most annotated to metabolism. An interesting find of this work is that the phyllosphere microbiota responsible for the synthesis of primary and secondary metabolites in A. argyi were significantly increased under a haze environment. Relatively enriched genes annotated by eggNOG belong to replication, recombination and repair, and genes classified into the glycoside hydrolase and glycosyltransferase enzymes were significantly increased. In summary, we found that both structure and function of phyllosphere microbiota are globally impacted by haze, while primary and secondary metabolites responsible for haze tolerance were considerably increased. These results suggest an adaptive strategy of plants for tolerating and confronting haze damage.

Genomic Analysis of Halotolerant Bacterial Strains Martelella soudanensis NC18T and NC20

  • Jung-Yun Lee;Dong-Hun Kim
    • Journal of Microbiology and Biotechnology
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    • 제32권11호
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    • pp.1427-1434
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    • 2022
  • Two novel, halotolerant strains of Martelella soudanensis, NC18T and NC20, were isolated from deep subsurface sediment, deeply sequenced, and comparatively analyzed with related strains. Based on a phylogenetic analysis using 16S rRNA gene sequences, the two strains grouped with members of the genus Martelella. Here, we sequenced the complete genomes of NC18T and NC20 to understand the mechanisms of their halotolerance. The genome sizes and G+C content of the strains were 6.1 Mb and 61.8 mol%, respectively. Moreover, NC18T and NC20 were predicted to contain 5,849 and 5,830 genes, and 5,502 and 5,585 protein-coding genes, respectively. Both strains contain the identically predicted 6 rRNAs and 48 tRNAs. The harboring of halotolerant-associated genes revealed that strains NC18T and NC20 might tolerate high salinity through the accumulation of potassium ions in a "salt-in" strategy induced by K+ uptake protein (kup) and the K+ transport system (trkAH and kdpFABC). These two strains also use the ectoine transport system (dctPQM), the glycine betaine transport system (proVWX), and glycine betaine uptake protein (opu) to accumulate "compatible solutes," such as ectoine and glycine betaine, to protect cells from salt stress. This study reveals the halotolerance mechanism of strains NC18T and NC20 in high salt environments and suggests potential applications for these halotolerant and halophilic strains in environmental biotechnology.

Transcriptome Analysis of Antrodia cinnamomea Mycelia from Different Wood Substrates

  • Jiao-Jiao Chen;Zhang Zhang;Yi Wang;Xiao-Long Yuan;Juan Wang;Yu-Ming Yang;Yuan Zheng
    • Mycobiology
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    • 제51권1호
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    • pp.49-59
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    • 2023
  • Antrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.