• Title/Summary/Keyword: Discovery DNA

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Distinctive Characteristics of an Autonomous Replication Sequence of Cephalosporium acremoniurn in Yeast (Cephalosporium acremonium의 자율복제 기점의 특성)

  • Lee, Kyoung;Kang, Dae-Wook;Yoon, Byung-Dae;Hwang, In-Kyu;Ahn, Jong-Seog;Min, Tae-Ick
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.215-221
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    • 1991
  • An autonomous replication sequence (ARS) derived from Cephalosporium acremonium ATCC 20339 was cloned in Sarchuromyces cerevisiae SHY 3 using YIp5 as a cloning vector. A new recombinant plasmid, designated pCY-2, which contained a 3.7 kb BamHI fragment of C. acrenzonium DNA showed the highest stability among the 40 recombinant plasmids composed of the YIp5 2nd ARS of C. ucremoniztm. Also, Southern hybridization and transformation of E, cull with DNA purified from yeast transformants verified that pCY-2 autonomously replicates in yeasts. Transformation efficiency and plasmid stability of pCY-2 in yeast were higher than those ol YRp 7 containing ARS which originated from yeast. Detailed studies by subcloning revealed that two ARSs existed within 2.6 kb of the insert, which is a novel discovery. However, it was concluded that these two ARSs were ligated during the gene manipulation in vitro.

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Pathway Analysis of Metabolic Syndrome Using a Genome-Wide Association Study of Korea Associated Resource (KARE) Cohorts

  • Shim, Unjin;Kim, Han-Na;Sung, Yeon-Ah;Kim, Hyung-Lae
    • Genomics & Informatics
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    • v.12 no.4
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    • pp.195-202
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    • 2014
  • Metabolic syndrome (MetS) is a complex disorder related to insulin resistance, obesity, and inflammation. Genetic and environmental factors also contribute to the development of MetS, and through genome-wide association studies (GWASs), important susceptibility loci have been identified. However, GWASs focus more on individual single-nucleotide polymorphisms (SNPs), explaining only a small portion of genetic heritability. To overcome this limitation, pathway analyses are being applied to GWAS datasets. The aim of this study is to elucidate the biological pathways involved in the pathogenesis of MetS through pathway analysis. Cohort data from the Korea Associated Resource (KARE) was used for analysis, which include 8,842 individuals (age, $52.2{\pm}8.9years$ ; body mass index, $24.6{\pm}3.2kg/m^2$). A total of 312,121 autosomal SNPs were obtained after quality control. Pathway analysis was conducted using Meta-analysis Gene-Set Enrichment of Variant Associations (MAGENTA) to discover the biological pathways associated with MetS. In the discovery phase, SNPs from chromosome 12, including rs11066280, rs2074356, and rs12229654, were associated with MetS (p < $5{\times}10^{-6}$), and rs11066280 satisfied the Bonferroni-corrected cutoff (unadjusted p < $1.38{\times}10^{-7}$, Bonferroni-adjusted p < 0.05). Through pathway analysis, biological pathways, including electron carrier activity, signaling by platelet-derived growth factor (PDGF), the mitogen-activated protein kinase kinase kinase cascade, PDGF binding, peroxisome proliferator-activated receptor (PPAR) signaling, and DNA repair, were associated with MetS. Through pathway analysis of MetS, pathways related with PDGF, mitogen-activated protein kinase, and PPAR signaling, as well as nucleic acid binding, protein secretion, and DNA repair, were identified. Further studies will be needed to clarify the genetic pathogenesis leading to MetS.

Profile Analysis of Bacteria in Human Hands Using the Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis (제한효소 절편길이 다형성(T-RFLP) 분석기법을 이용한 손에 서식하는 세균의 군집조성 분석)

  • Park, Jisun;Kim, Seung Bum
    • Journal of Science Criminal Investigation
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    • v.11 no.4
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    • pp.276-282
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    • 2017
  • As evidence that can be obtained at the crime scene, the importance of micro-evidences has been recognized in recent years with the development modern molecular-level analytical techniques. These micro-evidences include substances useful for personal identification such as DNA, but it is difficult to collect only the evidences showing individual characteristics every time at the crime scene. Therefore, development of new research approaches for the discovery and application of micro-evidence candidates is in increasing demand. For this purpose, skin microbial communities of bacteria inhabiting the palms of 16 people were collected and terminal-restriction enzyme fragment length polymorphism (T-RFLP) analysis was carried out to examine the potential for the application in personal identification. As a result, 16 different electropherograms were obtained, and various taxa including Staphylococcus and Bacillus were shown to produce different T-RF profiles among individuals. These results were analyzed with the factors affecting the microbiota such as sex and working environment of individuals.

Discovery of novel haplotypes from wild populations of Kappaphycus (Gigartinales, Rhodophyta) in the Philippines

  • Roleda, Michael Y.;Aguinaldo, Zae-Zae A.;Crisostomo, Bea A.;Hinaloc, Lourie Ann R.;Projimo, Vicenta Z.;Dumilag, Richard V.;Lluisma, Arturo O.
    • ALGAE
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    • v.36 no.1
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    • pp.1-12
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    • 2021
  • As the global demand for the carrageenophyte Kappaphycus is steadily increasing, its overall productivity, carrageenan quality, and disease resistance are gradually declining. In the face of this dilemma, wild Kappaphycus populations are viewed as sources of new cultivars that could potentially enhance production; therefore, assessment of their diversity is crucial. This study highlights the morphological and genetic diversity of wild Kappaphycus species obtained from two sites in the Philippines. Nucleotide alignments of available 5' region of the mitochondrial cytochrome c oxidase subunit I (COI-5P) and cox2-3 spacer sequences of Kappaphycus confirmed the presence of K. alvarezii in Guiuan, Eastern Samar and K. striatus in Bolinao, Pangasinan. Based on the concatenated sequences of the COI-5P and the cox2-3 spacer, nine novel haplotypes were observed along with other published haplotypes. However, there was no relationship between haplotype and morphology. These newly recognized haplotypes indicate a reservoir of unutilized wild genotypes in the Philippines, which could be taken advantage of in developing new cultivars with superior traits. DNA barcodes generated from this study effectively expand the existing databank of Kappaphycus sequences and can provide insights in elucidating the genetic diversity of Kappaphycus species in the country.

Five Alexandrium species lacking mixotrophic ability

  • Lim, An Suk;Jeong, Hae Jin;Ok, Jin Hee
    • ALGAE
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    • v.34 no.4
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    • pp.289-301
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    • 2019
  • Mixotrophy in marine organisms is an important aspect of ecology and evolution. The discovery of mixotrophic abilities in phototrophic dinoflagellates alters our understanding of the dynamics of red tides. In the phototrophic dinoflagellate genus Alexandrium, some species are mixotrophic, but others are exclusively autotrophic. There are differences in the ecological roles of autotrophic and mixotrophic Alexandrium in marine food webs. However, of the 34 known Alexandrium species, the mixotrophic ability of >20 species has yet to be explored. In this study, the mixotrophic capabilities of Alexandrium insuetum CCMP2082, Alexandrium mediterraneum CCMP3433, Alexandrium pacificum CCMP3434, Alexandrium tamutum ATSH1609, and Alexandrium margalefii CAWD10 were investigated by providing each species with 22 diverse prey items including bacterium-sized microbeads (1 ㎛), the cyanobacterium Synechococcus sp., algal prey species, and the ciliate Mesodinium rubrum. None of the 5 Alexandrium species fed on any of the prey items. These results increase the number of Alexandrium species lacking mixotrophic abilities to 9, compared to the 7 known mixotrophic Alexandrium species. Furthermore, the Alexandrium phylogenetic tree based on the large subunit ribosomal DNA contained 3 large clades, each of which had species with and without mixotrophic abilities. Thus, the acquisition or loss of mixotrophic abilities in Alexandrium might readily occur.

Opportunities and Challenges in Nutrigenomics and Health Promotion

  • Milner John A.
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2004.11a
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    • pp.17-23
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    • 2004
  • Not all individuals respond identically, or at times in the same direction, to dietary interventions. These inconsistencies likely arise because of diet and genomic interactions (nutrigenomics effects). A host of factors may influence the response to bioactive food components including specific polymorphisms (nutrigenetic effect), DNA methylation patterns and other epigenomic factors (nutritional epigenomic effects), capacity to induce anuo. suppress specific mRNA expression and patterns (nutritional transcriptomics), the occurrence and activity of proteins (proteomic effects), and/or the dose and temporal changes in cellular small molecular weight compounds will not only provide clues about specificity in response to food components, but assist in the identification of surrogate tissues and biomarkers that can predict a response. While this 'discovery' phase is critical for defining mechanisms and targets, and thus those who will benefit most from intervention, its true usefulness depends on moving this understanding into 'development' (interventions for better prevention, detection, diagnosis, and treatment) and a 'delivery' phase where information is provided to those most in need. It is incumbent on those involved with food and nutrition to embrace the 'omics' that relate to nutrition when considering not only the nutritional value of foods and their food components, but also when addressing acceptability and safety. The future of 'Nutrigenomics and Health Promotion' depends on the ability of the scientific community to identity appropriate biomarkers and susceptibility variants, effective communications about the merits of such undertakings with the health care community and with consumers, and doing all of this within a responsible bioethical framework.

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Structural Studies of Membrane Protein by Solid-state NMR Spectroscopy (고체상 핵자기공명 분광법을 이용한 막단백질의 구조연구)

  • Kim, Yongae
    • Analytical Science and Technology
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    • v.17 no.5
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    • pp.388-392
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    • 2004
  • Structural studies of membrane proteins, importantly involving interpretation of genomics information, many signaling pathway and major drug target for drug discovery, are having difficulty in characterizing the function using conventional solution nmr spectroscopy and x-ray crystallography because phospholipid bilayers hindered fast tumbling and crystallization. Here, we studied the structure of the pf1 coat protein in oriented phospholipid bilayers by home-built solid-state NMR probe. Bacteriophage pf1 was purified from Paeudomonas Aeruginosa and coat protein of bacteriophage pf1 was isolated from DNA and other proteins.

Characterization of Single Nucleotide Polymorphisms in 55 Disease-Associated Genes in a Korean Population

  • Lee, Seung-Ku;Kim, Hyoun-Geun;Kang, Jason-J.;Oh, Won-Il;Oh, Berm-Seok;Kwack, Kyu-Bum
    • Genomics & Informatics
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    • v.5 no.4
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    • pp.152-160
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    • 2007
  • Most common diseases are caused by multiple genetic and environmental factors. Among the genetic factors, single nucleotide polymorphisms (SNPs) are common DNA sequence variations in individuals and can serve as important genetic markers. Recently, investigations of gene-based and whole genome-based SNPs have been applied to association studies for marker discovery. However, SNPs are so population-specific that the association needs to be verified. Fifty-five genes and 384 SNPs were selected based on association with disease. Genotypes of 337 SNPs in candidate genes were determined using Illumina Sentrix Array Matrix (SAM) chips by an allele-specific extension method in 364 unrelated Korean individuals. Allelic frequencies of SNPs were compared with those of other populations obtained from the International HapMap database. Minor allele frequencies, linkage disequilibrium blocks, tagSNPs, and haplotypes of functional candidate SNPs in 55 genetic disease-associated genes were provided. Our data may provide useful information for the selection of genetic markers for gene-based genetic disease-association studies of the Korean population.

Genetic Risk Prediction for Normal-Karyotype Acute Myeloid Leukemia Using Whole-Exome Sequencing

  • Heo, Seong Gu;Hong, Eun Pyo;Park, Ji Wan
    • Genomics & Informatics
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    • v.11 no.1
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    • pp.46-51
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    • 2013
  • Normal-karyotype acute myeloid leukemia (NK-AML) is a highly malignant and cytogenetically heterogeneous hematologic cancer. We searched for somatic mutations from 10 pairs of tumor and normal cells by using a highly efficient and reliable analysis workflow for whole-exome sequencing data and performed association tests between the NK-AML and somatic mutations. We identified 21 nonsynonymous single nucleotide variants (SNVs) located in a coding region of 18 genes. Among them, the SNVs of three leukemia-related genes (MUC4, CNTNAP2, and GNAS) reported in previous studies were replicated in this study. We conducted stepwise genetic risk score (GRS) models composed of the NK-AML susceptible variants and evaluated the prediction accuracy of each GRS model by computing the area under the receiver operating characteristic curve (AUC). The GRS model that was composed of five SNVs (rs75156964, rs56213454, rs6604516, rs10888338, and rs2443878) showed 100% prediction accuracy, and the combined effect of the three reported genes was validated in the current study (AUC, 0.98; 95% confidence interval, 0.92 to 1.00). Further study with large sample sizes is warranted to validate the combined effect of these somatic point mutations, and the discovery of novel markers may provide an opportunity to develop novel diagnostic and therapeutic targets for NK-AML.