• Title/Summary/Keyword: NGS data analysis

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Draft Genome of Toxocara canis, a Pathogen Responsible for Visceral Larva Migrans

  • Kong, Jinhwa;Won, Jungim;Yoon, Jeehee;Lee, UnJoo;Kim, Jong-Il;Huh, Sun
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.751-758
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    • 2016
  • This study aimed at constructing a draft genome of the adult female worm Toxocara canis using next-generation sequencing (NGS) and de novo assembly, as well as to find new genes after annotation using functional genomics tools. Using an NGS machine, we produced DNA read data of T. canis. The de novo assembly of the read data was performed using SOAPdenovo. RNA read data were assembled using Trinity. Structural annotation, homology search, functional annotation, classification of protein domains, and KEGG pathway analysis were carried out. Besides them, recently developed tools such as MAKER, PASA, Evidence Modeler, and Blast2GO were used. The scaffold DNA was obtained, the N50 was 108,950 bp, and the overall length was 341,776,187 bp. The N50 of the transcriptome was 940 bp, and its length was 53,046,952 bp. The GC content of the entire genome was 39.3%. The total number of genes was 20,178, and the total number of protein sequences was 22,358. Of the 22,358 protein sequences, 4,992 were newly observed in T. canis. Following proteins previously unknown were found: E3 ubiquitin-protein ligase cbl-b and antigen T-cell receptor, zeta chain for T-cell and B-cell regulation; endoprotease bli-4 for cuticle metabolism; mucin 12Ea and polymorphic mucin variant C6/1/40r2.1 for mucin production; tropomodulin-family protein and ryanodine receptor calcium release channels for muscle movement. We were able to find new hypothetical polypeptides sequences unique to T. canis, and the findings of this study are capable of serving as a basis for extending our biological understanding of T. canis.

Plastome Phylogenomics of Commelinaceae Mirb. (Commelinales): Insights into Genome Evolution and Phylogenetic Relationships

  • Joonhyung Jung;Joo-Hwan Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.69-69
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    • 2022
  • Commelinaceae (Commelinales), consist of three subfamiles and 40 genera, are distributed in the Old and New world, except Europe. This family is commonly known as dayflower and spiderwort due to their short bloom time and a viscous stem secretion. Although, several morphological and molecular analysis were conducted, the relationships among the genera are still ambiguous. The rapid advances in next-generation sequencing (NGS) enable us to do genomic research widely. Here, we assembled 12 new plastomes of Commelinaceae including Cartonematoideae and compared with previously published data. We identified pseudogened accD and rpoA in Commelinoideae taxa. Phylogenetic analysis inferred from 78 protein-coding genes showed that Rhopalephora scaberrima was nested within Aneilema. Also, there is a need to revise the subtribal relationships in Tradescantieae. This study will contribute to define the genome structures, phylogenetic and biogeographic studies of Commelinaceae.

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Effective microbial molecular diagnosis of periodontitis-related pathogen Porphyromonas gingivalis from salivary samples using rgpA gene

  • Jinuk Jeong;Yunseok Oh;Junhyeon Jeon;Dong-Heon Baek;Dong Hee Kim;Kornsorn Srikulnath;Kyudong Han
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.13.1-13.8
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    • 2023
  • Importance of accurate molecular diagnosis and quantification of particular disease-related pathogenic microorganisms is highlighted as an introductory step to prevent and care for diseases. In this study, we designed a primer/probe set for quantitative real-time polymerase chain reaction (qRT-PCR) targeting rgpA gene, known as the specific virulence factor of periodontitis-related pathogenic bacteria 'Porphyromonas gingivalis', and evaluated its diagnostic efficiency by detecting and quantifying relative bacterial load of P. gingivalis within saliva samples collected from clinical subjects. As a result of qRT-PCR, we confirmed that relative bacterial load of P. gingivalis was detected and quantified within all samples of positive control and periodontitis groups. On the contrary, negative results were confirmed in both negative control and healthy groups. Additionally, as a result of comparison with next-generation sequencing (NGS)-based 16S metagenome profiling data, we confirmed relative bacterial load of P. gingivalis, which was not identified on bacterial classification table created through 16S microbiome analysis, in qRT-PCR results. It showed that an approach to quantifying specific microorganisms by applying qRT-PCR method could solve microbial misclassification issues at species level of an NGS-based 16S microbiome study. In this respect, we suggest that P. gingivalis-specific primer/probe set introduced in present study has efficient applicability in various oral healthcare industries, including periodontitis-related microbial molecular diagnosis field.

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.

Bioinformatics Resources of the Korean Bioinformation Center (KOBIC)

  • Lee, Byung-Wook;Chu, In-Sun;Kim, Nam-Shin;Lee, Jin-Hyuk;Kim, Seon-Yong;Kim, Wan-Kyu;Lee, Sang-Hyuk
    • Genomics & Informatics
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    • v.8 no.4
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    • pp.165-169
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    • 2010
  • The Korean Bioinformation Center (KOBIC) is a national bioinformatics research center in Korea. We developed many bioinformatics algorithms and applications to facilitate the biological interpretation of OMICS data. Here we present an introduction to major bioinformatics resources of databases and tools developed at KOBIC. These resources are classified into three main fields: genome, proteome, and literature. In the genomic resources, we constructed several pipelines for next generation sequencing (NGS) data processing and developed analysis algorithms and web-based database servers including miRGator, ESTpass, and CleanEST. We also built integrated databases and servers for microarray expression data such as MDCDP. As for the proteome data, VnD database, WDAC, Localizome, and CHARMM_HM web servers are available for various purposes. We constructed IntoPub server and Patome database in the literature field. We continue constructing and maintaining the bioinformatics infrastructure and developing algorithms.

Development of SNP markers for the identification of apple flesh color based on RNA-Seq data (RNA-Seq data를 이용한 사과 과육색 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Park, Seo Jun;Cho, Kang Hee;Lee, Han Chan;Lee, Jung Woo;Choi, In Myung
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.372-378
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    • 2017
  • For comparison of the transcription profiles in apple (Malus domestica L.) cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red flesh apple cultivar, 'Redfield' and white flesh apple cultivar, 'Granny Smith' were investigated by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the red flesh apple cultivar and white flesh apple cultivar were selected for nucleotide sequence determination and homology searches. High resolution melting (HRM) technique measures temperature induced strand separation of short PCR amplicons, and is able to detect variation as small as one base difference between red flesh apple cultivars and white flesh apple cultivars. We applied high resolution melting (HRM) analysis to discover single nucleotide polymorphisms (SNP) based on the predicted SNP information derived from the apple EST database. All 103 pairs of SNPs were discriminated, and the HRM profiles of amplicons were established. Putative SNPs were screened from the apple EST contigs by HRM analysis displayed specific difference between 10 red flesh apple cultivars and 11 white flesh apple cultivars. In this study, we report an efficient method to develop SNP markers from an EST database with HRM analysis in apple. These SNP markers could be useful for apple marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in apple cultivars.

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

  • Byun, Eun-Kyu;Kwak, Jae-Hyuck;Mun, Jihyeob
    • KIPS Transactions on Computer and Communication Systems
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    • v.8 no.10
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    • pp.231-238
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    • 2019
  • Analyzing next-generation genome sequencing data in a conventional way using single server may take several tens of hours depending on the data size. However, in order to cope with emergency situations where the results need to be known within a few hours, it is required to improve the performance of a single genome analysis. In this paper, we propose a parallelized method for pre-processing genome sequence data which can reduce the analysis time by utilizing the big data technology and the highperformance computing cluster which is connected to the high-speed network and shares the parallel file system. For the reliability of analytical data, we have chosen a strategy to parallelize the existing analytical tools and algorithms to the new environment. Parallelized processing, data distribution, and parallel merging techniques have been developed and performance improvements have been confirmed through experiments.

Comparative Chloroplast Analysis and Phylogenetic Relationships Among Corylopsis Siebold & Zucc. (Hamamelidaceae)

  • Tae-Hee Kim;Sang-Chul Kim;Young-Ho Ha;Hiroaki Setoguchi;Hyuk-Jin Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.55-55
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    • 2022
  • Corylopsis Siebold & Zucc. (Hamamelidaceae) is widely used for horticultural plant and comprise ca. 25 species in East Asia (1 species in Korea; 4 species in Japan; 20 species in China). Previous revisions have gone from 7 to more than 30 species, causing confusion in the nursery industry and public gardens. Due to morphological similarity within Corylopsis, molecular research is needed to distinguish it. In this study, the chloroplast genome of C. gotoana and C. pauciflora distributed in Japan was completed by using NGS (Next-Generation Sequencing) technique. The genome size of C. gotoana and C. pauciflora were 159,434 bp (large single-copy (LSC): 88,164 bp; small single-copy (SSC): 18,702 bp; inverted repeat regions (IRs): 26,284 bp) and 159,363 bp (LSC: 88,097 bp; SSC: 18,700 bp; IRs: 26,283 bp), respectively. In addition, we investigated the repeats, SNPs, and indels, and that could be used as DNA markers. Phylogenetic analysis demonstrated that C. pauciflora was sister to C. gotoana and C. spicata. The genus Corylopsis is a monophyletic group and Loropetalum is closely related to Corylopsis. The results of our study will provide the basic data necessary for the analysis of the species identification markers and genetic diversity within the genus Corylopsis in the future.

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Genetic Analysis Strategies for Improving Race Performance of Thoroughbred Racehorse and Jeju Horse (서러브레드 경주마와 제주마의 경주 능력 향상을 위한 유전체 분석 전략)

  • Baek, Kyung-Wan;Gim, Jeong-An;Park, Jung-Jun
    • Journal of Life Science
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    • v.28 no.1
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    • pp.130-139
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    • 2018
  • In ancient times, horse racing was done in ancient European countries in the form of wagon races or mountain races, and wagon racing was adopted as a regular event at the Greek Olympic Games. Thoroughbred horse has been bred since 17th century by intensive selective breeding for its speed, stamina, and racing ability. Then, in the 18th century, horse racing using the Thoroughbred species began to gain popularity among nobles. Since then, horse racing has developed into various forms in various countries and have developed into flat racing, steeplechasing, and harness racing. Thoroughbred racehorse has excellent racing abilities because of powerful selection breeding strategy for 300 years. It is necessary to maintain and maximize horses' ability to race, because horse industries produce enormous economic benefits through breeding, training, and horse racing. Next-generation sequencing (NGS) methods which process large amounts of genomic data have been developed recently. Based on the remarkable development of these genomic analytical techniques, it is now possible to easily carry out animal breeding strategies with superior traits. In order to select breeding racehorse with superior racing traits, the latest genomic analysis techniques have to be introduced. In this paper, we will review the current efforts to improve race performance for racehorses and to examine the research trends of genomic analysis. Finally, we suggest to utilize genomic analysis in Thoroughbred racehorse and Jeju horse, and propose a strategy for selective breeding for Jeju horse, which contributes job creation of Korea.

Comparative chloroplast genomics and phylogenetic analysis of the Viburnum dilatatum complex (Adoxaceae) in Korea

  • PARK, Jongsun;XI, Hong;OH, Sang-Hun
    • Korean Journal of Plant Taxonomy
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    • v.50 no.1
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    • pp.8-16
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    • 2020
  • Complete chloroplast genome sequences provide detailed information about any structural changes of the genome, instances of phylogenetic reconstruction, and molecular markers for fine-scale analyses. Recent developments of next-generation sequencing (NGS) tools have led to the rapid accumulation of genomic data, especially data pertaining to chloroplasts. Short reads deposited in public databases such as the Sequence Read Archive of the NCBI are open resources, and the corresponding chloroplast genomes are yet to be completed. The V. dilatatum complex in Korea consists of four morphologically similar species: V. dilatatum, V. erosum, V. japonicum, and V. wrightii. Previous molecular phylogenetic analyses based on several DNA regions did not resolve the relationship at the species level. In order to examine the level of variation of the chloroplast genome in the V. dilatatum complex, raw reads of V. dilatatum deposited in the NCBI database were used to reconstruct the whole chloroplast genome, with these results compared to the genomes of V. erosum, V. japonicum, and three other species in Viburnum. These comparative genomics results found no significant structural changes in Viburnum. The degree of interspecific variation among the species in the V. dilatatum complex is very low, suggesting that the species of the complex may have been differentiated recently. The species of the V. dilatatum complex share large unique deletions, providing evidence of close relationships among the species. A phylogenetic analysis of the entire genome of the Viburnum showed that V. dilatatum is a sister to one of two accessions of V. erosum, making V. erosum paraphyletic. Given that the overall degree of variation among the species in the V. dilatatum complex is low, the chloroplast genome may not provide a phylogenetic signal pertaining to relationships among the species.