• Title/Summary/Keyword: Gene organization

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A Gene Encoding Phosphatidyl Inositol-specific Phospholipase C form Cryphonectria parasitica Modulates the Hypoviral-modulated Laccase1 Expression

  • Kim, Dae-Hyuk
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2005.05a
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    • pp.159-161
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    • 2005
  • Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica is a useful model system to study the hypoviral regulation of fungal gene expression. The hypovirus is known to downregulate the fungal laccase1 (lac 1), the modulation of which is tightly governed by the inositol triphosphate ($IP_3$) and calcium second messenger system in a virus-free strain. We cloned the gene cplc1 encoding a phosphatidyl inositol-specific phospholipase C (PLC), in order to better characterize the fungal gene regulation by hypovirus. Sequence analysis of the cplc1 gene indicated that the protein product contained both the X and Y domains, which are the two conserved regions found in all known PLCs, with a 133 amino acid extension between the 2nd ${\beta}$-strand and the ${\alpha}$-helix in the X domain. In addition, the gene organization appeared to be highly similar to that of a ${\delta}$ type PLC. Disruption of the cplc1 gene resulted in slow growth and produced colonies characterized by little aerial mycelia and deep orange in color. In addition, down regulation of lac1 expression was observed. However, temperature sensitivity, osmosensitivity, virulence, and other hypovirulence-associated characteristics did not differ from the wild-type strain. Functional complementation of the cplc1-null mutant with the PLC1 gene from Saccharomyces cerevisiae restored lac1 expression, which suggests that the cloned gene encodes PLC activity. The present study indicates that the cplc1 gene is required for appropriate mycelial growth, and that it regulates the lac1 expression, which is also modulated by the hypovirus. Although several PLC genes have been identified in various simple eukaryotic organisms, the deletion analysis of the cplc1 gene in this study appears to be the first report on the functional analysis of PLC in filamentous fungi.

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Detection of hydin Gene Duplication in Personal Genome Sequence Data

  • Kim, Jong-Il;Ju, Young-Seok;Kim, Shee-Hyun;Hong, Dong-Wan;Seo, Jeong-Sun
    • Genomics & Informatics
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    • v.7 no.3
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    • pp.159-162
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    • 2009
  • Human personal genome sequencing can be done with high efficiency by aligning a huge number of short reads derived from various next generation sequencing (NGS) technologies to the reference genome sequence. One of the major obstacles is the incompleteness of human reference genome. We tried to analyze the effect of hidden gene duplication on the NGS data using the known example of hydin gene. Hydin2, a duplicated copy of hydin on chromosome 16q22, has been recently found to be localized to chromosome 1q21, and is not included in the current version of standard human genome reference. We found that all of eight personal genome data published so far do not contain hydin2, and there is large number of nsSNPs in hydin. The heterozygosity of those nsSNPs was significantly higher than expected. The sequence coverage depth in hydin gene was about two fold of average depth. We believe that these unique finding of hydin can be used as useful indicators to discover new hidden multiplication in human genome.

Genomic Structure and Phylogenetic Analysis of Luciferase Gene from the Firefly, Luciola lateralis

  • Kim, Jong-Gill;Park, Yong-Soo;Kim, Keun-Young;Bae, Jin-Sik;Kim, Iksoo;Park, Young-Cheol;Park, Ji-Young;Sohn, Hung-Dae;Jin, Byung-Rae
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.136-137
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    • 2003
  • Fireflies, the luminescent insect, have species specific flash patterns, being recognized as sexual communication. The lucifrrase gene is sole enzyme responsible for bioluminescence. The firefly luciferase gene is widely used as a genetic marker or as a reporter gene in a variety of organism including bacteria, plants and animals. In this study, we illustrate the complete organization of the genomic structure of the luciferase gene from L. lateralis sampled in Boun and Muju, Korea. (omitted)

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MediScore: MEDLINE-based Interactive Scoring of Gene and Disease Associations

  • Cho, Hye-Young;Oh, Bermseok;Lee, Jong-Keuk;Kim, Kuchan;Koh, InSong
    • Genomics & Informatics
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    • v.2 no.3
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    • pp.131-133
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    • 2004
  • MediScore is an information retrieval system, which helps to search for the set of genes associated with a specific disease or the set of diseases associated with a specific gene. Despite recent improvement of natural language processing (NLP) and other text mining approaches to search for disease associated genes, many false positive results come out due to diversity of exceptional cases as well as ambiguities in gene names. In order to overcome the weak points of current text mining approaches, MediScore introduces statistical normalization based on binomial to normal distribution approximation which corrects inaccurate scores caused by common words not representing genes and interactive rescoring by the user to remove the false positive results. Interactive rescoring includes individual alias scoring for each gene to remove false gene synonyms, referring MEDLINE abstracts, and cross referencing between OMIM and other related information.

Analysis of a Prodigiosin Biosynthetic Gene Cluster from the Marine Bacterium Hahella chejuensis KCTC 2396

  • Kim, Doc-Kyu;Park, Yon-Kyoung;Lee, Jong-Suk;F. Kim, Ji-Hyun;Jeong, Hae-Young;Kim, Beom-Seok;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1912-1918
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    • 2006
  • Marine bacterium Hahella chejuensis KCTC 2396 simultaneously produced red antibiotic prodigiosin and undecylprodiginine. A complete set of the prodigiosin biosynthetic gene cluster has been cloned, sequenced, and successfully expressed in a heterologous host. Sequence analysis of the gene cluster revealed 14 ORFs showing high similarity to pig and red genes from Serratia spp. and Streptomyces coelicolor A3(2), respectively, and the gene organization was almost: similar to that of pig genes. These genes were named hap for Hahella prodigiosin, and determined to be transcribed as a single operon, by RT-PCR experiment. Based on the hap gene mutagenesis experiments and comparative analysis with pig and red genes, we propose a prodigiosin-biosynthetic pathway in KCTC 2396.

Isolation and Analysis of the argG Gene Encoding Argininosuccinate Synthetase from Corynebacterium glutamicum

  • Ko, Soon-Young;Kim, Sei-Hyun;Lee, Heung-Shick;Lee, Myeong-Sok
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.949-954
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    • 2003
  • The argG gene of Corynebacterium glutamicum encoding argininosuccinate synthetase (EC6345) was cloned and sequenced. The gene was cloned by heterologous complementation of an Escherichia coli arginine auxotrophic mutant (argG/sup -/). The cloned DNA fragment also complements E. coli argD, argF, and argH mutants, suggesting a clustered organization of the genes in the chromosome. The coding region of the argG gene is 1,206 nucleotides long with a deduced molecular weight of about 44 kDa, comparable with the predicted size of the expressed protein on the SDS-PAGE. Computer analysis revealed that the amino acid sequence of the argG gene product had a high similarity to that of Mycobacterium tuberculosis and Streptomyces clavuligerus. Two conserved sequence motifs within the ArgG appear to be ATP-binding sites which correspond to 2 of the 3 conserved regions found in sequences of all known argininosuccinate synthetases.

Identification of the Phenalamide Biosynthetic Gene Cluster in Myxococcus stipitatus DSM 14675

  • Park, Suhyun;Hyun, Hyesook;Lee, Jong Suk;Cho, Kyungyun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1636-1642
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    • 2016
  • Phenalamide is a bioactive secondary metabolite produced by Myxococcus stipitatus. We identified a 56 kb phenalamide biosynthetic gene cluster from M. stipitatus DSM 14675 by genomic sequence analysis and mutational analysis. The cluster is comprised of 12 genes (MYSTI_04318- MYSTI_04329) encoding three pyruvate dehydrogenase subunits, eight polyketide synthase modules, a non-ribosomal peptide synthase module, a hypothetical protein, and a putative flavin adenine dinucleotide-binding protein. Disruption of the MYSTI_04324 or MYSTI_04325 genes by plasmid insertion resulted in a defect in phenalamide production. The organization of the phenalamide biosynthetic modules encoded by the fifth to tenth genes (MYSTI_04320-MYSTI_04325) was very similar to that of the myxalamid biosynthetic gene cluster from Stigmatella aurantiaca Sg a15, as expected from similar backbone structures of the two substances. However, the loading module and the first extension module of the phenalamide synthase encoded by the first to fourth genes (MYSTI_04326-MYSTI_04329) were found only in the phenalamide biosynthetic gene cluster from M. stipitatus DSM 14675.

Molecular Characterization of Rockbream (Oplegnathus fasciatus) Cytoskeletal β-actin Gene and Its 5'-Upstream Regulatory Region

  • Lee, Sang-Yoon;Kim, Ki-Hong;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.12 no.2
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    • pp.90-97
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    • 2009
  • The cytoskeletal $\beta$-actin gene and its 5'-upstream region were isolated and characterized in the rockbream (Oplegnathus fasciatus). Complementary DNA of the rockbream $\beta$-actin represented a 1,125 bp of an open reading frame encoding 375 amino acids, and the rockbream $\beta$-actin cDNA and deduced amino acid sequences were highly homologous to those of other vertebrate orthologs. At the genomic level, the $\beta$-actin gene also exhibited an organization typical of vertebrate cytoskeletal actin genes (2,159 bp composed of five translated exons interrupted by four introns) with a conserved GT/AG exon-intron splicing rule. The putative non-translated exon predicted in the rockbream $\beta$-actin gene was much more homologous with those of teleostean $\beta$-actin genes than those of mammals. The 5'-upstream regulatory region isolated by genome walking displayed conserved and essential elements such as TATA, CArG and CAAT boxes in its proximal part, while several other immune- or stress-related motifs such as those for NF-kappa B, USF, HNF, AP-1 and C/EBP were in the distal part. Semi-quantitative RT-PCR assay results demonstrated that the rockbream $\beta$-actin transcripts were ubiquitously but different-tially expressed across the tissues of juveniles.