• Title/Summary/Keyword: Shotgun

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Reduction of Ambiguity in Phosphorylation-site Localization in Large-scale Phosphopeptide Profiling by Data Filter using Unique Mass Class Information

  • Madar, Inamul Hasan;Back, Seunghoon;Mun, Dong-Gi;Kim, Hokeun;Jung, Jae Hun;Kim, Kwang Pyo;Lee, Sang-Won
    • Bulletin of the Korean Chemical Society
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    • v.35 no.3
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    • pp.845-850
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    • 2014
  • The rapid development of shotgun proteomics is paving the way for extensive proteome profiling, while providing extensive information on various post translational modifications (PTMs) that occur to a proteome of interest. For example, the current phosphoproteomic methods can yield more than 10,000 phosphopeptides identified from a proteome sample. Despite these developments, it remains a challenging issue to pinpoint the true phosphorylation sites, especially when multiple sites are possible for phosphorylation in the peptides. We developed the Phospho-UMC filter, which is a simple method of localizing the site of phosphorylation using unique mass classes (UMCs) information to differentiate phosphopeptides with different phosphorylation sites and increase the confidence in phosphorylation site localization. The method was applied to large scale phosphopeptide profiling data and was demonstrated to be effective in the reducing ambiguity associated with the tandem mass spectrometric data analysis of phosphopeptides.

Analysis of sequence alignment Tools on polymorphic genomes (다염기변이 유전체에 대한 서열 정렬 툴 분석)

  • Kim, Yoo-Sun;Kim, Jong-Hyun;Yeo, Yun-Ku;Kim, Woo-Cheol;Park, Sang-Hyun
    • Proceedings of the Korean Information Science Society Conference
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    • 2008.06c
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    • pp.217-221
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    • 2008
  • 생명공학 기술의 발달로 지놈 프로젝트를 통해 인간 초파리 등 여러 종의 유전체 정보가 밝혀 졌다. 그러나 Post-Genome 연구에 있어서 매우 중요한 생물체인 멍게(Ciona intestinalis)와 성게(Strongylocentrotus purpuratus)의 유전체 서열은 현재 공개되어 있으나 염기서열의 연속성(continuity)에는 심각한 문제점이 존재하고 있다. 이들은 염기서열에 변이가 많은 다염기변이 유전체(polymorphic genomes)로 그 특성이 반영되지 않은 전통적인 Whole Genome Shotgun Sequencing(WGSS)방법을 사용였기 때문이다. 이와 같은 다염기변이 유전체 서열 분석은 시스템 생물학이나 비교 유전체학 등의 후발 연구에 기초가 되므로 매우 중요하다. 본 논문에서는 다염기변이 유전체에 대해 알아보고 서열 조립 알고리즘의 기본이 되는 서열 정렬 툴들 중 가장 많이 사용되는 FASTA, BLAST, BLAT에 대해 분석하여 봄으로써 다염기변이 유전체에 적합한 서열 조립 전략 수립을 위해 고려해야 하는 사항들을 논의해 본다.

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Inference of Aspergillus fumigatus Pathways by Computational Genome Analysis: Tricarboxylic Acid Cycle (TCA) and Glyoxylate Shunt

  • Do, Jin-Hwan;Anderson, Michael-J.;Denning, David-W.;Erich, Bornberg-Bauer
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.74-80
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    • 2004
  • Aspergillus fumigatus is one of the most common fungi in the human environment, both in-doors and out-doors. It is the main causative agent of invasive aspergillosis, a life-threatening mycosis among immunocompromised patients. The genome has been sequenced by an international consortium, including the Wellcome Trust Sanger Institute (U.K.) and The Institute for Genomic Research (TIGR, U.S.A.), and a ten times whole genome shotgun sequence assembly has been made publicly available. In this study, we identified tricarboxylic acid (TCA) cycle enzymes of A. fumigatus by comparative analysis with four other fungal species. The open reading frames showed high amino acid sequence similarity with the other fungal citric acid enzymes and well-conserved functional domains. All genes present in Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans, and Neurospora crassa were also found in A. fumigatus. In addition, we identified four A. fumigatus genes coding for enzymes in the glyoxylate shunt, which may be required for fungal virulence. The architecture of multi-gene encoded enzymes, such as isocitrate dehydrogenase, 2-ketoglutarate, succinyl-CoA synthetase, and succinate dehydrogenase was well conserved in A. fumigatus. Furthermore, our results show that genes of A. fumigatus can be detected reliably using GlimmerM.

A Study on Immersive Audio Improvement of FTV using an effective noise (유효 잡음을 활용한 FTV 입체음향 개선방안 연구)

  • Kim, Jong-Un;Cho, Hyun-Seok;Lee, Yoon-Bae;Yeo, Sung-Dae;Kim, Seong-Kweon
    • The Journal of the Korea institute of electronic communication sciences
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    • v.10 no.2
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    • pp.233-238
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    • 2015
  • In this paper, we proposed that immersive audio effect method using the effective noise to improve engagement in free-viewpoint TV(FTV) service. In the basketball court, we monitored the frequency spectrums by acquiring continuous audio data of players and referee using shotgun and wireless microphone. By analyzing this spectrum, in case that users zoomed in, we determined whether it is effective frequency or not. Therefore when users using FTV service zoom in toward the object, it is proposed that we need to utilize unnecessary noise instead of removing that. it will be able to be useful for an immersive audio implementation of FTV.

Identification of Antibiotic Resistance Genes in Orofacial Abscesses Using a Metagenomics-based Approach: A Pilot Study

  • Yeeun Lee;Joo-Young Park;Youngnim Choi
    • Journal of Korean Dental Science
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    • v.16 no.1
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    • pp.35-46
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    • 2023
  • Purpose: Culture-based methods for microbiological diagnosis and antibiotic susceptibility tests have limitations in the management of orofacial infections. We aimed to profile pus microbiota and identify antibiotic resistance genes (ARGs) using a culture-independent approach. Materials and Methods: Genomic DNA samples extracted from the pus specimens of two patients with orofacial abscesses were subjected to shotgun sequencing on the NovaSeq system. Taxonomic profiling and prediction of ARGs were performed directly from the metagenomic raw reads. Result: Taxonomic profiling revealed obligate anaerobic polymicrobial communities associated with infections of odontogenic origins: the microbial community of Patient 1 consisted of one predominant species (Prevotella oris 74.6%) with 27 minor species, while the sample from Patient 2 contained 3 abundant species (Porphyromonas endodontalis 33.0%; P. oris 31.6%; and Prevotella koreensis 13.4%) with five minor species. A total of 150 and 136 putative ARGs were predicted in the metagenome of each pus sample. The coverage of most predicted ARGs was less than 10%, and only the CfxA2 gene identified in Patient 1 was covered 100%. ARG analysis of the seven assembled genome/metagenome datasets of P. oris revealed that strain C735 carried the CfxA2 gene. Conclusion: A metagenomics-based approach is useful to profile predominantly anaerobic polymicrobial communities but needs further verification for reliable ARG detection.

Metagenome Analysis of Protein Domain Collocation within Cellulase Genes of Goat Rumen Microbes

  • Lim, SooYeon;Seo, Jaehyun;Choi, Hyunbong;Yoon, Duhak;Nam, Jungrye;Kim, Heebal;Cho, Seoae;Chang, Jongsoo
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.8
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    • pp.1144-1151
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    • 2013
  • In this study, protein domains with cellulase activity in goat rumen microbes were investigated using metagenomic and bioinformatic analyses. After the complete genome of goat rumen microbes was obtained using a shotgun sequencing method, 217,892,109 pair reads were filtered, including only those with 70% identity, 100-bp matches, and thresholds below $E^{-10}$ using METAIDBA. These filtered contigs were assembled and annotated using blastN against the NCBI nucleotide database. As a result, a microbial community structure with 1431 species was analyzed, among which Prevotella ruminicola 23 bacteria and Butyrivibrio proteoclasticus B316 were the dominant groups. In parallel, 201 sequences related with cellulase activities (EC.3.2.1.4) were obtained through blast searches using the enzyme.dat file provided by the NCBI database. After translating the nucleotide sequence into a protein sequence using Interproscan, 28 protein domains with cellulase activity were identified using the HMMER package with threshold E values below $10^{-5}$. Cellulase activity protein domain profiling showed that the major protein domains such as lipase GDSL, cellulase, and Glyco hydro 10 were present in bacterial species with strong cellulase activities. Furthermore, correlation plots clearly displayed the strong positive correlation between some protein domain groups, which was indicative of microbial adaption in the goat rumen based on feeding habits. This is the first metagenomic analysis of cellulase activity protein domains using bioinformatics from the goat rumen.

Heavy Metal Concentrations of Some Game Species Captured in Kyeongsangnam-do, Korea (경남지역(慶南地域)에 서식(棲息)하는 수렵조수류(狩獵鳥獸類)의 중금속(重金屬) 축적(蓄積))

  • Choi, Jai Sik;Jung, Wee Hyun;Youn, Ki Sick;Lee, Doo Pyo
    • Journal of Korean Society of Forest Science
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    • v.83 no.1
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    • pp.6-11
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    • 1994
  • Three different types of tissue taken from three game birds such as Ring-necked Pheasant, Rufous Turtle Dove, Mallard and a mammal, Korean Water Deer were analyzed for five heavy metals such as iron, zinc, copper, lead and cadmium. The main purpose of this study was to make clear the background levels of the metal concentrations in the tissues of these game species. The metal levels were generally low and could be considered as the background levels. But some markedly high levels of lead were recorded in the muscles of birds shot by the shotgun. The implications of these results were discussed with respect to the health of people.

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Cloning of Bacillus amyloliquefaciens amylase gene using YRp7 as a vector I. Expression of cloned amylase gene in Escherichia coli (YRp 7 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I. Escherichia coli에서의 발현)

  • 서정훈;김영호;전도연;홍순덕;조윤래
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.161-168
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    • 1986
  • A 1.95Kb Sau3Al fragment coding for $\alpha$-amylase from Bacillus amyloliquefaciens was isolated by the shotgun method using Escherichia coli as a host. The genome of Bacillus amyloliquefaciens was partially digested with the restriction endonuclease Sau3Al and joined to plasmid YRp7 cleaved with the restriction endonuclease BamHI. The $\alpha$-amylase gene present in a 1.95Kb insert was stably maintained and expressed in Escherichia coli. The amount of $\alpha$-amylase activity produced by Escherichia coli containing the hybrid plasmid pEA24 was about 65% of the activity produced by the donor Bacillus amyloliquefaciens strain. The properties of $\alpha$-amylase produced by Escherichia coli were very similar to those produced by Bacillus amyloliquefaciens as based on optimum temperature, pH, and effect of CaCl$_2$ concentration. About 70% of the $\alpha$-amylase produced by Escherichia coli was localized in the periplasmic space, whereas the remaining enzyme was localized in the inner part of the cell.

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Cloning of Bacillus amyloliquefaciens amylase gene using YEp13 as a vector I. Expression of cloned amylase gene in Escherichia coli (YEp 13 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I. Escherichia coli에서의 발현)

  • 이창후;서정훈
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.155-160
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    • 1986
  • $\alpha$-Amylase gene of B. amyloliquefaciens was cloned to E. coli-yeast shuttle vector YEp-13 and expressed in E. coli. Chromosomal DNA of B. amyloliquefaciens was partially digested with Sau3Al and YEp13 plasmid was cleaved with BamH1. The hybrid plasmid, pHA28, was constructed by shotgun method and transformed to E. coli C600 and HB101. The amount of $\alpha$-amylase produced by transformants of E. coli was about 20% to 30% of that produced by B. amyloli-quefaciens. About 65% of $\alpha$-amylase produced by transformant was secreted into periplasm and the others were located in cytoplasm. $\alpha$-Amylase production was maximal when transformants were cultivated for 15hr to 20hr. As the result of agarose gel electrophoresis, pHA28 plasmid was found to be various in its size. This result suggested that pHA28 plasmid was segregated.

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Comparative Whole Cell Proteomics of Listeria monocytogenes at Different Growth Temperatures

  • Won, Soyoon;Lee, Jeongmin;Kim, Jieun;Choi, Hyungseok;Kim, Jaehan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.259-270
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    • 2020
  • Listeria monocytogenes is a gram-positive, facultative anaerobe food pathogen responsible for the listeriosis that mostly occurs during the low-temperature storage of a cold cut or dairy products. To understand the systemic response to a wide range of growth temperatures, L. monocytogenes were cultivated at a different temperature from 10℃ to 42℃, then whole cell proteomic analysis has been performed both exponential and stationary cells. The specific growth rate increased proportionally with the increase in growth temperature. The maximum growth rate was observed at 37℃ and was maintained at 42℃. Global protein expression profiles mainly depended on the growth temperatures showing similar clusters between exponential and stationary phases. Expressed proteins were categorized by their belonging metabolic systems and then, evaluated the change of expression level in regard to the growth temperature and stages. DnaK, GroEL, GroES, GrpE, and CspB, which were the heat&cold shock response proteins, increased their expression with increasing the growth temperatures. In particular, GroES and CspB were expressed more than 100-fold than at low temperatures during the exponential phase. Meanwhile, CspL, another cold shock protein, overexpressed at a low temperature then exponentially decreased its expression to 65-folds. Chemotaxis protein CheV and flagella proteins were highly expressed at low temperatures and stationary phases. Housekeeping proteins maintained their expression levels constant regardless of growth temperature or growth phases. Most of the growth related proteins, which include central carbon catabolic enzymes, were highly expressed at 30℃ then decreased sharply at high growth temperatures.