• 제목/요약/키워드: Illumina MiSeq

검색결과 60건 처리시간 0.026초

Whole genome sequence of Staphylococcus aureus strain RMI-014804 isolated from pulmonary patient sputum via next-generation sequencing technology

  • Ayesha, Wisal;Asad Ullah;Waheed Anwar;Carlos M. Morel;Syed Shah Hassan
    • Genomics & Informatics
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    • 제21권3호
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    • pp.34.1-34.10
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    • 2023
  • Nosocomial infections, commonly referred to as healthcare-associated infections, are illnesses that patients get while hospitalized and are typically either not yet manifest or may develop. One of the most prevalent nosocomial diseases in hospitalized patients is pneumonia, among the leading causes of mortality and morbidity. Viral, bacterial, and fungal pathogens cause pneumonia. More severe introductions commonly included Staphylococcus aureus, which is at the top of bacterial infections, per World Health Organization reports. The staphylococci, S. aureus, strain RMI-014804, mesophile, on-sporulating, and non-motile bacterium, was isolated from the sputum of a pulmonary patient in Pakistan. Many characteristics of S. aureus strain RMI-014804 have been revealed in this paper, with complete genome sequence and annotation. Our findings indicate that the genome is a single circular 2.82 Mbp long genome with 1,962 protein-coding genes, 15 rRNA, 49 tRNA, 62 pseudogenes, and a GC content of 28.76%. As a result of this genome sequencing analysis, researchers will fully understand the genetic and molecular basis of the virulence of the S. aureus bacteria, which could help prevent the spread of nosocomial infections like pneumonia. Genome analysis of this strain was necessary to identify the specific genes and molecular mechanisms that contribute to its pathogenicity, antibiotic resistance, and genetic diversity, allowing for a more in-depth investigation of its pathogenesis to develop new treatments and preventive measures against infections caused by this bacterium.

유용한 바실러스의 토양 접종에 따른 토착 세균 군집의 변화 (Changes in Resident Soil Bacterial Communities in Response to Inoculation of Soil with Beneficial Bacillus spp.)

  • 김이슬;김상윤;안주희;상미경;원항연;송재경
    • 한국미생물·생명공학회지
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    • 제46권3호
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    • pp.253-260
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    • 2018
  • 유용미생물은 임업과 축산 분야에 활용될 뿐만 아니라 병해충 방제와 작물 생육 증진 등의 용도로 농업에서 널리 이용되고 있다. 하지만 유용미생물의 토양에서의 생존율과 정착율에 대한 연구는 미미한 형편이다. 본 연구에서는 마이크로코즘을 이용해 바실러스 3 균주를 토양에 처리한 후, 이들의 토양 내 생존능을 정량 PCR을 이용하여 13일 동안 정량적으로 분석하였다. 또한 Illumina MiSeq 플랫폼을 이용하여 바실러스 3 균주 처리구와 대조구의 토양미생물 군집 분포를 비교 및 분석하였다. 바실러스 3 균주의 처리 직후 토양 내 밀도는 건조토양 1 그람당 평균 $4.4{\times}10^6$ 유전자수로 대조구에 비해 1,000배 이상 높았다. 바실러스 균주의 토양 내 밀도는 처리 후 약 일주일 간 유지되었고 그 후부터는 유의성 있게 감소하였지만 여전히 대조구보다 100배 이상 높았다. 바실러스 균주 처리 후 토양 내 미생물 군집 구조 분석 결과, 대조구와 처리구 모두 Acidobacteria 문($26.3{\pm}0.9%$), Proteobacteria 문($24.2{\pm}0.5%$), Chloroflexi 문($11.1{\pm}0.4%$), Actinobacteria 문($9.7{\pm}2.5%$)에 속하는 세균이 우점하였다. 대조구 대비 처리구에서 Actinobacteria 문의 비율은 뚜렷하게 감소하였지만 Bacteroidetes 문과 Firmicutes 문의 비율은 증가하는 경향이었다. 속 수준에서 바실러스 3 균주를 처리함에 따라 일부 세균 군집의 종 풍부도를 변화되었고, 결국 전체 토착 미생물 군집 구조가 변화되었음을 확인할 수 있었다. 본 연구에서 수행한 유용한 바실러스의 토양 접종 후 이들의 토양 내 생존능 분석 및 토착 세균 군집의 변화는 유용미생물을 생물적 제제로 시설재배지에 사용할 때 중요한 정보를 제공할 것으로 판단된다.

Transcriptional Profiling of Differentially Expressed Genes in Porcine Satellite Cell

  • Jeong, Jin Young;Kim, Jang Mi;Rajesh, Ramanna Valmiki;Suresh, Sekar;Jang, Gul Won;Lee, Kyung-Tai;Kim, Tae Hun;Park, Mina;Jeong, Hak Jae;Kim, Kyung Woon;Cho, Yong Min;Lee, Hyun-Jeong
    • Reproductive and Developmental Biology
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    • 제37권4호
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    • pp.233-245
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    • 2013
  • Muscle satellite cell (SC) is responsible for postnatal muscle growth, repair, and regeneration. Satellite cell is an important source of multi-potent stem cell process and differentiation into adipogenic, myogenic, and osteoblastogenic. The objective of this study was to identify alter of transcriptome during differentiation in porcine satellite cell and to elevated transcriptome at different stages of postnatal development to gain insight into the differences in differentiated PSC. We used RNA-seq technique to investigate the transcriptomes during differentiation in pig muscle. Sequence reads were obtained from Illumina HiSeq2000. Differentially expressed genes (DEG) were detected by EdgeR. Gene ontology (GO) terms are powerful tool for unification among representation genes or products. In study of GO biological terms, functional annotation clustering involved in cell cycle, apoptosis, extracellular matrix, phosphorylation, proteolysis, and cell signaling in differences stage. Taken together, these results would be contributed to a better understanding of muscle biology and processes underlying differentiation. Our results suggest that the source of DEGs could be better understanding of the mechanism of muscle differentiation and transdifferentiation.

닭의 성숙/미성숙란에서 RNA Sequencing을 이용한 유전자 발현 양상 고찰 (Gene Expression Profiling by RNA Sequencing in Mature/Immature Oocytes of Chicken)

  • 강경수;장현준;박미나;최정우;정원형;허강녕;최창용;김영주;이시우;조은석;김남신;김태헌;한재용;이경태
    • 한국가금학회지
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    • 제41권4호
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    • pp.287-296
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    • 2014
  • 조류의 난포 성장은 호르몬의 작용에 따라 크기가 달라져 각각의 단계를 이루며 성장하게 된다. 난의 성숙에 관련된 유전자는 난 단백질 생산과 산란률에 밀접한 관련이 있으며, 이를 유전자 발현 측면에서 심도 있는 고찰이 필요가 있다. 본 연구는 NGS를 이용한 RNA-seq 데이터를 이용하여 유전자의 발현량과 유전자 상호 구조에 대한 분석을 실시하여 난의 발달 과정에 필요한 유전자군을 조사하였다. 본 실험에 사용된 개체는 한국 재래계 흑색계통이 사용되었고, 비교조직은 미성숙란과 성숙란의 RNA를 추출하여 유전자의 발현 양상을 살펴봄으로 난의 성숙에 필요한 유전자의 발현 양상을 보고자 하였다. 실험을 위해 Total RNA를 추출하였고, HiSeq 2000 platform을 사용하여 염기서열을 분석하고, Tuxedo Protocol과 DAVID 프로그램을 통해 유전자의 기능과 상호간의 연관관계를 예측하였다. 탐색된 유전자군은 미성숙란과 성숙란 간에 많은 차이를 보이고 있는 유전자군을 탐색한 결과, 315개의 발현이 다르게 나타나는 것으로 보이고 있으며, GO 분석을 통하여 기능면에서 미성숙란과 성숙란에서 확연히 구분되는 유전자 발현 양상을 확인할 수 있었다. 이들 결과를 통하여 향후 난성숙 과정을 이해하고, 계란 품질 향상을 위한 마커 개발을 기여할 수 있을 것으로 사료된다.

Differential Impacts on Bacterial Composition and Abundance in Rhizosphere Compartments between Al-Tolerant and Al-Sensitive Soybean Genotypes in Acidic Soil

  • Wen, Zhong-Ling;Yang, Min-Kai;Fazal, Aliya;Liao, Yong-Hui;Cheng, Lin-Run;Hua, Xiao-Mei;Hu, Dong-Qing;Shi, Ji-Sen;Yang, Rong-Wu;Lu, Gui-Hua;Qi, Jin-Liang;Hong, Zhi;Qian, Qiu-Ping;Yang, Yong-Hua
    • Journal of Microbiology and Biotechnology
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    • 제30권8호
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    • pp.1169-1179
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    • 2020
  • In this study, two soybean genotypes, i.e., aluminum-tolerant Baxi 10 (BX10) and aluminumsensitive Bendi 2 (BD2), were used as plant materials and acidic red soil was used as growth medium. The soil layers from the inside to the outside of the root are: rhizospheric soil after washing (WRH), rhizospheric soil after brushing (BRH) and rhizospheric soil at two sides (SRH), respectively. The rhizosphere bacterial communities were analyzed by high-throughput sequencing of V4 hypervariable regions of 16S rRNA gene amplicons via Illumina MiSeq. The results of alpha diversity analysis showed that the BRH and SRH of BX10 were significantly lower in community richness than that of BD2, while the WRH exhibited no significant difference between BX10 and BD2. Among the three sampling compartments of the same soybean genotype, WRH had the lowest community richness and diversity while showing the highest coverage. Beta diversity analysis results displayed no significant difference for any compartment between the two genotypes, or among the three different sampling compartments for any same soybean genotype. However, the relative abundance of major bacterial taxa, specifically nitrogen-fixing and/or aluminum-tolerant bacteria, was significantly different in the compartments of the BRH and/or SRH at phylum and genus levels, indicating genotype-dependent variations in rhizosphere bacterial communities. Strikingly, as compared with BRH and SRH, the WRH within the same genotype (BX10 or BD2) always had an enrichment effect on rhizosphere bacteria associated with nitrogen fixation.

Fine mapping of rice bacterial leaf blight resistance loci to major Korean races of Xoo (Xanthomonas oryzae)

  • Lee, Myung-Chul;Choi, Yu-Mi;Lee, Sukyeung;Yoon, Hyemyeong;Oh, Sejong
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 추계학술대회
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    • pp.73-73
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    • 2018
  • Bacterial leaf blight(BLB), caused by X. oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice due to its high epidemic potential. Understanding BLB resistance at a genetic level is important to further improve the rice breeding that provides one of the best approaches to control BLB disease. In the present investigation, a collection of 192 accessions was used in the genome-wide association study (GWAS) for BLB resistance loci against four Korean races of Xoo that were represented by the prevailing BLB isolates under Xoo differential system. A total of 192 accessions of rice germplasm were selected on the basis of the bioassay using four isolated races of Xoo such as K1, K2, K3 and K3a. The selected accessions was used to prepare 384-plex genotyping by sequencing (GBS) libraries and Illumina HiSeq 2000 paired- end read was used for GBS sequencing. GWAS was conducted using T ASSEL 5.0. The T ASSEL program uses a mixed linear model (MLM). T he results of the bioassay using a selected set of 192 accessions showed that a large number of accessions (93.75%) were resistant to K1 race, while the least number of accessions (34.37%) resisted K3a race. For races K2 and K3, the resistant germplasm proportion remained between 66.67 to 70.83%. T he genotypic data produced SNP matrix for a total of 293,379 SNPs. After imputation the missing data was removed, which exhibited 34,724 SNPs for association analysis. GWAS results showed strong signals of association at a threshold of [-log10(P-value)] more than5 (K1 and K2) and more than4 (K3 and K3a) for nine of the 39 SNPs, which are plausible candidate loci of resistance genes. T hese SNP loci were positioned on rice chromosome 2, 9, and 11 for K1 and K2 races, whereas on chromosome 4, 6, 11, and 12 for K3 and K3a races. The significant loci detected have also been illustrated, NBS-LRR type disease resistance protein, SNARE domain containing protein, Histone deacetylase 19, NADP-dependent oxidoreductase, and other expressed and unknown proteins. Our results provide a better understanding of the distribution of genetic variation of BLB resistance to Korean pathogen races and breeding of resistant rice.

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토마토에 염류 내성을 유도하는 바실러스 균주 처리 후 근권 미생물 군집 구조 연구 (Assessment of Rhizosphere Microbial Community Structure in Tomato Plants after Inoculation of Bacillus Species for Inducing Tolerance to Salinity)

  • 유성제;이신애;원항연;송재경;상미경
    • 한국환경농학회지
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    • 제40권1호
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    • pp.49-59
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    • 2021
  • BACKGROUND: Soil salinity causes reduction of crop productivity. Rhizosphere microbes have metabolic capabilities and ability to adaptation of plants to biotic and abiotic stresses. Plant growth-promoting bacteria (PGPB) could play a role as elicitors for inducing tolerance to stresses in plants by affecting resident microorganism in soil. This study was conducted to demonstrate the effect of selected strains on rhizosphere microbial community under salinity stress. METHODS AND RESULTS: The experiments were conducted in tomato plants in pots containing field soil. Bacterial suspension was inoculated into three-week-old tomato plants, one week after inoculation, and -1,000 kPa-balanced salinity stress was imposed. The physiological and biochemical attributes of plant under salt stress were monitored by evaluating pigment, malondialdehyde (MDA), proline, soil pH, electrical conductivity (EC) and ion concentrations. To demonstrate the effect of selected Bacillus strains on rhizosphere microbial community, soil microbial diversity and abundance were evaluated with Illumina MiSeq sequencing, and primer sets of 341F/805R and ITS3/ITS4 were used for bacterial and fungal communities, respectively. As a result, when the bacterial strains were inoculated and then salinity stress was imposed, the inoculation decreases the stress susceptibility including reduction in lipid peroxidation, enhanced pigmentation and proline accumulation which subsequently resulted in better plant growth. However, bacterial inoculations did not affect diversity (observed OTUs, ACE, Chao1 and Shannon) and structure (principle coordinate analysis) of microbial communities under salinity stress. Furthermore, relative abundance in microbial communities had no significant difference between bacterial treated- and untreated-soils under salinity stress. CONCLUSION: Inoculation of Bacillus strains could affect plant responses and soil pH of tomato plants under salinity stress, whereas microbial diversity and abundance had no significant difference by the bacterial treatments. These findings demonstrated that Bacillus strains could alleviate plant's salinity damages by regulating pigments, proline, and MDA contents without significant changes of microbial community in tomato plants, and can be used as effective biostimulators against salinity stress for sustainable agriculture.

Genome characterization and mutation analysis of human influenza A virus in Thailand

  • Rattanaburi, Somruthai;Sawaswong, Vorthon;Nimsamer, Pattaraporn;Mayuramart, Oraphan;Sivapornnukul, Pavaret;Khamwut, Ariya;Chanchaem, Prangwalai;Kongnomnan, Kritsada;Suntronwong, Nungruthai;Poovorawan, Yong;Payungporn, Sunchai
    • Genomics & Informatics
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    • 제20권2호
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    • pp.21.1-21.14
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    • 2022
  • The influenza A viruses have high mutation rates and cause a serious health problem worldwide. Therefore, this study focused on genome characterization of the viruses isolated from Thai patients based on the next-generation sequencing technology. The nasal swabs were collected from patients with influenza-like illness in Thailand during 2017-2018. Then, the influenza A viruses were detected by reverse transcription-quantitative polymerase chain reaction and isolated by MDCK cells. The viral genomes were amplified and sequenced by Illumina MiSeq platform. Whole genome sequences were used for characterization, phylogenetic construction, mutation analysis and nucleotide diversity of the viruses. The result revealed that 90 samples were positive for the viruses including 44 of A/H1N1 and 46 of A/H3N2. Among these, 43 samples were successfully isolated and then the viral genomes of 25 samples were completely amplified. Finally, 17 whole genomes of the viruses (A/H1N1, n=12 and A/H3N2, n=5) were successfully sequenced with an average of 232,578 mapped reads and 1,720 genome coverage per sample. Phylogenetic analysis demonstrated that the A/H1N1 viruses were distinguishable from the recommended vaccine strains. However, the A/H3N2 viruses from this study were closely related to the recommended vaccine strains. The nonsynonymous mutations were found in all genes of both viruses, especially in hemagglutinin (HA) and neuraminidase (NA) genes. The nucleotide diversity analysis revealed negative selection in the PB1, PA, HA, and NA genes of the A/H1N1 viruses. High-throughput data in this study allow for genetic characterization of circulating influenza viruses which would be crucial for preparation against pandemic and epidemic outbreaks in the future.

Whole-genome resequencing reveals domestication and signatures of selection in Ujimqin, Sunit, and Wu Ranke Mongolian sheep breeds

  • Wang, Hanning;Zhong, Liang;Dong, Yanbing;Meng, Lingbo;Ji, Cheng;Luo, Hui;Fu, Mengrong;Qi, Zhi;Mi, Lan
    • Animal Bioscience
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    • 제35권9호
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    • pp.1303-1313
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    • 2022
  • Objective: The current study aimed to perform whole-genome resequencing of Chinese indigenous Mongolian sheep breeds including Ujimqin, Sunit, and Wu Ranke sheep breeds (UJMQ, SNT, WRK) and deeply analyze genetic variation, population structure, domestication, and selection for domestication traits among these Mongolian sheep breeds. Methods: Blood samples were collected from a total of 60 individuals comprising 20 WRK, 20 UJMQ, and 20 SNT. For genome sequencing, about 1.5 ㎍ of genomic DNA was used for library construction with an insert size of about 350 bp. Pair-end sequencing were performed on Illumina NovaSeq platform, with the read length of 150 bp at each end. We then investigated the domestication and signatures of selection in these sheep breeds. Results: According to the population and demographic analyses, WRK and SNT populations were very similar, which were different from UJMQ populations. Genome wide association study identified 468 and 779 significant loci from SNT vs UJMQ, and UJMQ vs WRK, respectively. However, only 3 loci were identified from SNT vs WRK. Genomic comparison and selective sweep analysis among these sheep breeds suggested that genes associated with regulation of secretion, metabolic pathways including estrogen metabolism and amino acid metabolism, and neuron development have undergone strong selection during domestication. Conclusion: Our findings will facilitate the understanding of Chinese indigenous Mongolian sheep breeds domestication and selection for complex traits and provide a valuable genomic resource for future studies of sheep and other domestic animal breeding.

저지종 젖소의 반추위 내 미생물 균총 변화와 고창증 발병간의 상관관계 연구 (The Study on the Relationship between Changes of Rumen Microflora and Bloat in Jersey Cow)

  • 김상범;오종석;정하연;정영훈;박범영;하승민;임석기;이성실;박지후;박성민;김언태
    • 한국초지조사료학회지
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    • 제38권2호
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    • pp.106-111
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    • 2018
  • 본 연구는 고창증 발병으로 폐사된 저지종 젖소의 반추위액과 캐놀라가 장착한 저지종 젖소의 반추위액을 샘플링하여 반추위 내 미생물 균총의 급작스런 변화에 따른 반추위 발효와 고창증 발병간에 연관성을 알아보고자 진행되었다. 채취한 반추위액 샘플은 PowerMax Soil DNA Isolation Kit (MO BIO Inc., CA, USA)를 이용하여 DNA를 추출한 후 PicoGreen (Turner BioSystems, Inc., CA, USA)과 Nanodrop spectrophotometer (ND-1000, NanoDrop Technologies Inc., DE, USA)를 이용하여 260/280 nm와 260/230 nm 흡광도 값을 측정하였다. DNA를 정량분석하고 $MiSeq^{TM}$ platform (Illumina, CA, USA)을 이용한 장내 미생물 균총의 다양성 분석을 마크로젠(Macrogen Inc., Seoul)에 의뢰하여 실시하였다. 반추위액 내 미생물 균총 분석 결과, 전분 분해균의 경우 고창증 발병으로 인한 폐사우의 반추위 내에서 R. bromii가 우점 하였으며, R. bromii, B. pseudolongum, B. merycicum 및 B. fibrisolvens 등과 같은 전분 분해균들이 정상우 대비 36배 높았다. 반면 반추위 발효와 밀접한 관련이 있는 F. succinogenes, R. albus 및 R. flavefaciens 등과 같은 대표적인 섬유소 분해균 비율은 정상우에서 고창증 발병으로 인한 폐사우 대비 12배 높았다. 이와 같은 결과를 볼 때, 반추위 내 균총 가운데 R. bromii, B. pseudolongum, B. merycicum 및 B. fibrisolvens 등과 같은 전분 분해균들의 급격한 증가가 반추위 내 pH 저하 및 가스 생성 증가를 초래했고 이는 저지종 젖소의 고창증 발병으로 인한 폐사와 밀접한 연관이 있었을 것으로 판단된다.