• Title/Summary/Keyword: Model sequencing

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Lessons from the Sea : Genome Sequence of an Algicidal Marine Bacterium Hahella chehuensis (적조 살상 해양 미생물 Hahella chejuensis의 유전체 구조)

  • Jeong Hae-Young;Yoon Sung-Ho;Lee Hong-Kum;Oh Tae-Kwang;Kim Ji-Hyun
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.1-6
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    • 2006
  • Harmful algal blooms (HABs or red tides), caused by uncontrolled proliferation of marine phytoplankton, impose a severe environmental problem and occasionally threaten even public health. We sequenced the genome of an EPS-producing marine bacterium Hahella chejuensis that produces a red pigment with the lytic activity against red-tide dinoflagellates at parts per billion level. H. chejuensis is the first sequenced species among algicidal bacteria as well as in the order Oceanospirillales. Sequence analysis indicated a distant relationship to the Pseudomonas group. Its 7.2-megabase genome encodes basic metabolic functions and a large number of proteins involved in regulation or transport. One of the prominent features of the H. chejuensis genome is a multitude of genes of functional equivalence or of possible foreign origin. A significant proportion (${\sim}23%$) of the genome appears to be of foreign origin, i.e. genomic islands, which encode genes for biosynthesis of exopolysaccharides, toxins, polyketides or non-ribosomal peptides, iron utilization, motility, type III protein secretion and pigment production. Molecular structure of the algicidal pigment was determined to be prodigiosin by LC-ESI-MS/MS and NMR analyses. The genomics-based research on H. chejuensis opens a new possibility for controlling algal blooms by exploiting biotic interactions in the natural environment and provides a model in marine bioprospecting through genome research.

Bacillus licheniformis Isolated from Korean Traditional Food Sources Enhances the Resistance of Caenorhabditis elegans to Infection by Staphylococcus aureus

  • Yun, Hyun Sun;Heo, Ju Hee;Son, Seok Jun;Park, Mi Ri;Oh, Sangnam;Song, Min-Ho;Kim, Jong Nam;Go, Gwang-Woong;Cho, Ho-Seong;Choi, Nag-Jin;Jo, Seung-Wha;Jeong, Do-Youn;Kim, Younghoon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1105-1108
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    • 2014
  • We investigated whether Bacillus spp., newly isolated from Korean traditional food resources, influence the resistance of hosts to foodborne pathogens, by using Caenorhabditis elegans as a surrogate host model. Initially, we selected 20 Bacillus spp. that possess antimicrobial activity against various foodborne pathogens, including Staphylococcus aureus. Among the selected strains, six strains of Bacillus spp. used in preconditioning significantly prolonged the survival of nematodes exposed to S. aureus. Based on 16S rRNA gene sequencing, all six strains were identified as B. licheniformis. Our findings suggest that preconditioning with B. licheniformis may modulate the host defense response against S. aureus.

Evaluation of Biogas Production Performance and Archaeal Microbial Dynamics of Corn Straw during Anaerobic Co-Digestion with Cattle Manure Liquid

  • Zhang, Benyue;Zhao, Hongyan;Yu, Hairu;Chen, Di;Li, Xue;Wang, Weidong;Piao, Renzhe;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.739-747
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    • 2016
  • The rational utilization of crop straw as a raw material for natural gas production is of economic significance. In order to increase the efficiency of biogas production from agricultural straw, seasonal restrictions must be overcome. Therefore, the potential for biogas production via anaerobic straw digestion was assessed by exposing fresh, silage, and dry yellow corn straw to cow dung liquid extract as a nitrogen source. The characteristics of anaerobic corn straw digestion were comprehensively evaluated by measuring the pH, gas production, chemical oxygen demand, methane production, and volatile fatty acid content, as well as applying a modified Gompertz model and high-throughput sequencing technology to the resident microbial community. The efficiency of biogas production from fresh straw (433.8 ml/g) was higher than that of production from straw silage and dry yellow straw (46.55 ml/g and 68.75 ml/g, respectively). The cumulative biogas production from fresh straw, silage straw, and dry yellow straw was 365 l-1 g-1 VS, 322 l-1 g-1 VS, and 304 l-1 g-1 VS, respectively, whereas cumulative methane production was 1,426.33%, 1,351.35%, and 1,286.14%, respectively, and potential biogas production was 470.06 ml-1 g-1 VS, 461.73 ml-1 g-1 VS, and 451.76 ml-1 g-1 VS, respectively. Microbial community analysis showed that the corn straw was mainly metabolized by acetate-utilizing methanogens, with Methanosaeta as the dominant archaeal community. These findings provide important guidance to the biogas industry and farmers with respect to rational and efficient utilization of crop straw resources as material for biogas production.

Saccharomyces cerevisiae KNU5377 with Multiple Stress Tolerance and its Potential as a Worldwide On-site Industrial Strain for Alcohol Fermentation

  • Paik, Sang-Kyoo;Ingnyol Jin;Yun, Hae-Sun;Park, Sae-Hun;Shin, Seong-Chul;Kim, Jae-Wan;Shin, Ki-Sun;Lee, Jung-Sook;Park, Yong-Ha
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.425-429
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    • 2002
  • Saccharomyces cerevisiae KNU5377 was examined to assay the recovering capacity against heat and other stressors. Along with a particular fermentation ability that is able to produce ethanol even at high temperature such as $40^{\circ}C$ with a comparable rate to the fermentation at $33^{\circ}C$, this strain also exhibited higher viability than a reference strain owing to its own thermotolerance that conferred the survival after the severe heat shock at $60^{\circ}C$ for 30 minutes. Furthermore, this strain showed outstanding tolerances against $H_2O_2$, ethanol and some chemical compounds. But, especially due to the thermotolerance, this strain has been suspected of other species of yeast. However, ITS (internally transcribed spacer) 1 and 2 sequencing data confirmed this strain was a typical strain of S. cerevisiae. The outstanding tolerances to various environmental stressors Indicate this S. cerevisiae KNU5377 is enough to use both as an on-site potential strain for world-wide alcohol fermentation industry and as a model strain for researches into the routes to acquire the tolerance to various stressors.

Cloning, Characterization and Expression Analysis of Interleukin-10 from the Zebrafish (Danio rerion)

  • Zhang, Dian-Chang;Shao, Yan-Qing;Huang, Yan-Qin;Jiang, Shi-Gui
    • BMB Reports
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    • v.38 no.5
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    • pp.571-576
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    • 2005
  • Cytokines are proteins produced by many different cells of the immune system and play a significant role in initiating and regulating the inflammatory process. In this research, an important cytokine, interleukin-10 (IL-10) gene, has been identified and characterized from zebrafish (Danio rerio) genome database. Zebrafish IL-10 is located within a 2690 bp fragment and contains five exons and four introns, sharing the same organization with mammalian IL-10 genes. An open reading frame of 543 bp was found to encode a putative 180 amino acid protein with a signal peptide of 22 amino acids, which shares 29.7-80.9% homology with amino acid sequences of other known IL-10. The signature motif of IL-10 is also conserved in zebrafish IL-10. The predicted transcript was finally confirmed by sequencing of cDNA clones. Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the tissue distribution and expression regulation of this gene in seven organs of normal and lipopolysaccharide (LPS) stimulation zebrafish. The results demonstrated that this gene was expressed slightly in normal kidney, gill and gut, no expression was detected in other four tissues. The expression was clearly upregulated after LPS stimulation. Using the ideal zebrafish model, further study of IL-10 characterization and function may provide insight on the understanding of the innate immune system.

Subtype Distribution of Blastocystis in Communities along the Chao Phraya River, Thailand

  • Palasuwan, Attakorn;Palasuwan, Duangdao;Mahittikorn, Aongart;Chiabchalard, Rachatawan;Combes, Valery;Popruk, Supaluk
    • Parasites, Hosts and Diseases
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    • v.54 no.4
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    • pp.455-460
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    • 2016
  • Blastocystis is a common zoonotic enteric protozoan that has been classified into 17 distinct subtypes (STs). A cross-sectional study was conducted to determine the prevalence and subtype distributions of Blastocystis in villagers living along the Chao Phraya River, Ayutthaya Province, Thailand, and to assess the risk of zoonotic infection. In total, 220 stool samples were collected, and DNA was extracted. PCR and sequencing were performed with primers targeting the small-subunit ribosomal RNA (SSU rRNA) genes. Blastocystis was present in 5.9% (13/220) of samples, and ST3 (5.0%; 11/220) was the predominant subtype, followed by ST2 (0.45%; 1/220) and ST6 (0.45%; 1/220). Phylogenetic trees were constructed with the maximum-likelihood method based on the Hasegawa-Kishino-Yano + G + I model, neighbor-joining, and maximum parsimony methods. The percentage of bootstrapped trees in which the associated taxa clustered together was relatively high. All the sequences of the Blastocystis-positive samples (KU051524-KU051536) were closely related to those from animals (pig, cattle, and chicken), indicating a zoonotic risk. Therefore, the villagers require proper health education, especially regarding the prevention of parasitic infection, to improve their personal hygiene and community health. Further studies are required to investigate the Blastocystis STs in the animals living in these villages.

Association Analysis between Polymorph isms of NOTCH4 Gene and Schizophrenia in Korean Population

  • Lee, Seo-Kyong;Kang, Sung-Wook;Kim, Su-Kang;Kim, Hak-Jae;Kim, Tae;Park, Jin-Kyung;Cho, Ah-Rang;Kim, Jong-Woo;Park, Hyun-Kyung;Kim, Youn-Jung;Choe, Bong-Keun;Song, Ji-Young
    • Molecular & Cellular Toxicology
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    • v.5 no.2
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    • pp.160-164
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    • 2009
  • Notch signaling plays a crucial role in development of the nervous system. Neurodevelopmental hypothesis on etiology of schizophrenia has been implicated. The aim of this study is to determine whether single nucleotide polymorphisms (SNPs) of Notch homolog 4 (Drosophila) (NOTCH4) gene are associated with schizophrenia. This study included 283 schizophrenia patients diagnosed according to DSM-IV and 301 normal control subjects. Control subjects without history of psychiatric disorders were recruited. Four missense SNPs [rs915894 (exon 3, Lys117Gln), rs2071282 (exon 4, Pro204Leu), rs422951 (exon 6, Thr320Ala), and rs17604492 (exon 18, Gly942Arg)] of NOTCH4 gene were genotyped by the direct sequencing method. Multiple logistic regression models (codominant, dominant, and recessive models) were employed to evaluate odds ratio, 95% confidence interval, and p value. For analysis of genetic data, SNPStats, Haploview, HapAnalyzer, SNPAnalyzer, and Helixtree programs were also used. Of 4 SNPs, rs2071282 was weekly associated with schizophrenia in two alternative models (codominant model, P=0.049; dominant, P=0.041). However, these associations were not significant after Bonferroni correction. At 4 SNPs, one linkage disequilibrium (LD) block was made. This block consisted of rs915894 and rs2071282. In haplotype analysis, AC haplotype was weakly associated with schizophrenia (dominant, P=0.04). This association was disappeared after Bonferroni correction. Our result shows possibility that some SNPs of NOTCH4 gene may be weekly associated with development of schizophrenia in Korean population. However, replication result by other population will be needed.

BRCA1 Gene Mutation Screening for the Hereditary Breast and/or Ovarian Cancer Syndrome in Breast Cancer Cases: a First High Resolution DNA Melting Analysis in Indonesia

  • Mundhofir, Farmaditya EP;Wulandari, Catharina Endah;Prajoko, Yan Wisnu;Winarni, Tri Indah
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1539-1546
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    • 2016
  • Specific patterns of the hereditary breast and ovarian cancer (HBOC) syndrome are related to mutations in the BRCA1 gene. One hundred unrelated breast cancer patients were interviewed to obtain clinical symptoms and signs, pedigree and familial history of HBOC syndrome related cancer. Subsequently, data were calculated using the Breast and Ovarian Analysis of Disease Incidence and Carrier Estimation Algorithm (BOADICEA) risk prediction model. Patients with high score of BOADICEA were offered genetic testing. Eleven patients with high score of BOADICEA, 2 patients with low score of BOADICEA, 2 patient's family members and 15 controls underwent BRCA1 genetic testing. Mutation screening using PCR-HRM was carried out in 22 exons (41 amplicons) of BRCA1 gene. Sanger sequencing was subjected in all samples with aberrant graph. This study identified 10 variants in the BRCA1 gene, consisting of 6 missense mutations (c.1480C>A, c.2612C>T, c.2566T>C, c.3113A>G, c.3548 A>G, c.4837 A>G), 3 synonymous mutations (c.2082 C>T, c.2311 T>C and c.4308T>C) and one intronic mutation (c.134+35 G>T). All variants tend to be polymorphisms and unclassified variants. However, no known pathogenic mutations were found.

High-density genetic mapping using GBS in Chrysanthemum

  • Chung, Yong Suk;Cho, Jin Woong;Kim, Changsoo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.57-57
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    • 2017
  • Chrysanthemum is one of the most important floral crop in Korea produced about 7 billion dollars (1 billion for pot and 6 billion for cutting) in 2013. However, it is difficult to breed and to do genetic study because 1) it is highly self-incompatible, 2) it is outcrossing crop having heterozygotes, and 3) commercial cultvars are hexaploid (2n = 6x = 54). Although low-density genetic map and QTL study were reported, it is not enough to apply for the marker assisted selection and other genetic studies. Therefore, we are trying to make high-density genetic mapping using GBS with about 100 $F_1s$ of C. boreale that is oHohhfd diploid (2n = 2x = 18, about 2.8Gb) instead of commercial culitvars. Since Chrysanthemum is outcrossing, two-way pseudo-testcross model would be used to construct genetic map. Also, genotype-by-sequencing (GBS) would be utilized to generate sufficient number of markers and to maximize genomic representation in a cost effective manner. Those completed sequences would be analyzed with TASSEL-GBS pipeline. In order to reduce sequence error, only first 64 sequences, which have almost zero percent error, would be incorporated in the pipeline for the analysis. In addition, to reduce errors that is common in heterozygotes crops caused by low coverage, two rare cutters (NsiI and MseI) were used to increase sequence depth. Maskov algorithm would also used to deal with missing data. Further, sparsely placed markers on the physical map would be used as anchors to overcome problems caused by low coverage. For this purpose, were generated from transcriptome of Chrysanthemum using MISA program. Among those, 10 simple sequence repeat (SSR) markers, which are evenly distributed along each chromosome and polymorphic between two parents, would be selected.

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NICKEL INCORPORATION INTO Klebsiella aerogenes UREASE (Klebsiella aerogenes Urease로의 닉켈의 도입)

  • Lee, Mann-Hyung-
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.69-80
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    • 1994
  • Although ureases play important roles in microbial nitrogen metabolism and in the pathogenesis of several human diseases, little is known of the mechanism of metallocenter biosynthesis in this Ni-Containing enzyme. Klebsiella aerogenes urease apo-protein was purified from cells grown in the absence of Ni. The purified apo-enzyme showed the same native molecular weight, charge, and subunit stoichiometry as the holo-enzyme. Chemical modification studies were consistent with histidinyl ligation of Ni. Apo-enzyme could not be activated by simple addition of Ni ions suggesting a requirement for a cellular factor. Deletion analysis showed that four accessory genes (ureD, ureE, ureF, and ureG) are necessary for the functional incorporation of the urease metallocenter. Whereas the $\Delta$ureD, $\Delta$ureF, and $\Delta$ureG mutants are inactive and their ureases lack Ni, the $\Delta$ureE mutants retain partial activity and their ureases possess corresponding lower levels of Ni. UreE and UreG peptides were identified by SDS-polyacrylamide gel comparisons of mutant and wild type cells and by N-terminal sequencing. UreD and UreF peptides, which are synthesized at ve교 low levels, were identified by using in vitro transcription/translation methods. Cotransformation of E. coli cells with the complementing plasmids confirmed that ureD and ureF gene products act in trans. UreE was purified and characterized. immunogold electron microscopic studies were used to localize UreE to the cytoplasm. Equilibrium dialysis studies of purified UreE with $^{63}$ NiC1$_2$ showed that it binds ~6 Ni in a specific manner with a $K_{d}$ of 9.6 $\pm$1.3 $\mu$M. Results from spectroscopic studies demonstrated that Ni ions are ligated by 5 histidinyl residues and a sixth N or O atom, consistent with participation of the polyhistidine tail at the carboxyl termini of the dimeric UreE in Ni binding. With these results and other known features of the urease-related gene products, a model for urease metallocenter biosynthesis is proposed in which UreE binds Ni and acts as a Ni donor to the urease apo-protein while UreG binds ATP and couples its Hydrolysis to the Ni incorporation process.ouples its Hydrolysis to the Ni incorporation process.s.

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