• Title/Summary/Keyword: gene cluster

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Cloning, Sequencing, and Characterization of the Pradimicin Biosynthetic Gene Cluster of Actinomadura hibisca P157-2

  • Kim, Byung-Chul;Lee, Jung-Min;Ahn, Jong-Seog;Kim, Beom-Seok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.830-839
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    • 2007
  • Pradimicins are potent antifungal antibiotics having an unusual dihydrobenzo[$\alpha$]naphthacenequinone aglycone substituted with D-alanine and sugars. Pradimicins are polyketide antibiotics produced by Actinomadura hibisca P157-2. The gene cluster involved in the biosynthesis of pradimicins was cloned and sequenced. The pradimicin gene cluster was localized to a 39-kb DNA segment and its involvement in the biosynthesis of pradimicin was proven by gene inactivation of prmA and prmB(ketosynthases $\alpha\;and\;\beta$). The pradimicin gene cluster consists of 28 open reading frames(ORFs), encoding a type II polyketide synthase(PKS), the enzymes involved in sugar biosynthesis and tailoring enzymes as well as two resistance proteins. The deduced proteins showed strong similarities to the previously validated gene clusters of angucyclic polyketides such as rubromycin, griseorhodin, and fredericamycin. From the pradimicin gene cluster, prmP3 encoding a component of the acetyl-CoA carboxylase complex was disrupted. The production levels of pradimicins of the resulting mutants decreased to 62% of the level produced by the wild-type strain, which indicate that the acetyl-CoA carboxylase gene would have a significant role in the production of pradimicins through supplying the extender unit precursor, malonyl-CoA.

Comparison of the Cluster Validation Methods for High-dimensional (Gene Expression) Data (고차원 (유전자 발현) 자료에 대한 군집 타당성분석 기법의 성능 비교)

  • Jeong, Yun-Kyoung;Baek, Jang-Sun
    • The Korean Journal of Applied Statistics
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    • v.20 no.1
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    • pp.167-181
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    • 2007
  • Many clustering algorithms and cluster validation techniques for high-dimensional gene expression data have been suggested. The evaluations of these cluster validation techniques have, however, seldom been implemented. In this paper we compared various cluster validity indices for low-dimensional simulation data and real gene expression data, and found that Dunn's index is the most effective and robust, Silhouette index is next and Davies-Bouldin index is the bottom among the internal measures. Jaccard index is much more effective than Goodman-Kruskal index and adjusted Rand index among the external measures.

Comparison of the Cluster Validation Techniques using Gene Expression Data (유전자 발현 자료를 이용한 군집 타당성분석 기법 비교)

  • Jeong, Yun-Kyoung;Baek, Jang-Sun
    • 한국데이터정보과학회:학술대회논문집
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    • 2006.04a
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    • pp.63-76
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    • 2006
  • Several clustering algorithms to analyze gene expression data and cluster validation techniques that assess the quality of their outcomes, have been suggested, but evaluations of these cluster validation techniques have seldom been implemented. In this paper we compared various cluster validity indices for simulation data and real genomic data, and found that Dunn's index is more effective and robust through small simulations and with real gene expression data.

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New Gene Cluster from Thermophile Bacillus fordii MH602 for Conversion of DL-5-Substituted Hydantoins to L-Amino Acids

  • Mei, Yan-Zhen;Wan, Yong-Min;He, Bing-Fang;Ying, Han-Jie;Ouyang, Ping-Kai
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1497-1505
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    • 2009
  • The thermophile Bacillus fordii MH602 was screened for stereospecifically hydrolyzing DL-5-substituted hydantoins to L-$\alpha$-amino acids. Since the reaction occurs at higher temperature, the advantages for enhancement of substrate solubility and for racemization of DL-5-substituted hydantoins during the conversion were achieved. The hydantoin metabolism gene cluster from thermophile is firstly reported in this paper. The genes involved in hydantoin utilization (hyu) were isolated on an 8.2-kb DNA fragment by restriction site-dependent PCR, and six ORFs were identified by DNA sequence analysis. The hyu gene cluster contained four genes with novel cluster organization characteristics: the hydantoinase gene hyuH, putative transport protein gene hyuP, hyperprotein gene hyuHP, and L-carbamoylase gene hyuC. The hyuH and hyuC genes were heterogeneously expressed in E. coli. The results indicated that hyuH and hyuC are involved in the conversion of DL-5-substituted hydantoins to an N-carbamyl intermediate that is subsequently converted to L-$\alpha$-amino acids. Hydantoinase and carbamoylase from B. fordii MH602 compared respectively with reported hydantoinase and carbamoylase showed the highest identities of 71% and 39%. The novel cluster organization characteristics and the difference of the key enzymes between thermopile B. fordii MH602 and other mesophiles were presumed to be related to the evolutionary origins of concerned metabolism.

Molecular Structure of the PHA Synthesis Gene Cluster from New mcl-PHA Producer Pseudomonas putida KCTC1639

  • KIM TAE-KWON;VO MINH TRI;SHIN HYUN-DONG;LEE YONG-HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1120-1124
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    • 2005
  • Pseudomonas putida KCTC 1639 was newly identified as a potential producer of biodegradable medium chain length polyhydroxyalkanoates. It exhibited a carbon assimilation pattern quite different from other known P. putida strains, but a more similar pattern with P. oleovorans, which assimilates the carbon sources mainly through ${\beta}$-oxidation rather than the fatty acid biosynthesis pathway. The PHA synthesis gene cluster from P. putida KCTC1639 was composed of two gene loci; the PHA synthase gene locus and granule-associated gene locus, which were cloned and deposited in the GenBank under accession numbers AY286491 and AY750858 as a new nucleotide sequence, respectively. The molecular structure and amino acid homology of the new gene cluster were compared with those from Pseudomonas species, including other P. putida strains and P. oleovorans, and a higher than $90\%$ homology was observed.

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.

Cloning and Characterization of a Gene Cluster for the Production of Polyketide Macrolide Dihydrochalcomycin in Streptomyces sp. KCTC 0041BP

  • Jaishy Bharat Prasad;Lim Si-Kyu;Yoo Ick-Dong;Yoo Jin-Cheol;Sohng Jae-Kyung;Nam Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.764-770
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    • 2006
  • Dihydrochalcomycin (GERI-155), produced by Streptomyces sp. KCTC-0041BP isolated from Korean soil, is a 16-membered macrolide antibiotic consisting of two deoxysugar moieties at C-5 and C-20 positions of a branched lactone ring. The cloning and sequencing of a gene cluster for dihydrochalcomycin biosynthesis revealed a 63-kb nucleotide region containing 25 open reading frames (ORFs). The products of all of these 25 ORFs playa role in dihydrochalcomycin biosynthesis and self-resistance against the compounds synthesized. At the core of this cluster lies a 39.6-kb polyketide synthase (PKS) region encoding eight modules in five giant multifunctional protein-coding genes (gerSI-SV). The genes responsible for the biosynthesis of deoxysugar moieties, D-chalcose and D-mycinose, and their modification and attachment were found on either side of this PKS region. The involvement of this gene cluster in dihydrochalcomycin biosynthesis was confirmed by disruption of the dehydratase (DH) domain in module 3 of the PKS gene and by metabolite analysis.

Genome of Betaproteobacterium Caenimonas sp. Strain SL110 Contains a Coenzyme $F_{420}$ Biosynthesis Gene Cluster

  • Li, Xiuling;Feng, Fuying;Zeng, Yonghui
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1490-1494
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    • 2014
  • To probe the genomic properties of microbes thriving in desert lakes, we sequenced the full genome of a betaproteobacterial strain (SL110) belonging to the understudied genus Caenimonas of the family Comamonadaceae. This strain was isolated from a freshwater lake in the western Gobi Desert, Northern China. Its genome contains genes encoding carbon monoxide dehydrogenase, nitrate reductase, nitrite reductase, nitric oxide reductase, and sulfur oxidation enzymes, highlighting the potentially important contribution of this group of bacteria to the cycling of inorganic elements in nature. Unexpectedly, a coenzyme $F_{420}$ biosynthesis gene cluster was identified. A further search for $F_{420}$ biosynthesis gene homologs in genomic databases suggests the possible widespread presence of $F_{420}$ biosynthesis gene clusters in proteobacterial genomes.

The Chloroplast rpl23 Gene Cluster of Spirogyra maxima (Charophyceae) Shares Many Similarities with the Angiosperm rpl23 Operon

  • Lee, Jung-Ho;James R. Manhart
    • ALGAE
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    • v.17 no.1
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    • pp.59-68
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    • 2002
  • A phylogenetic affinity between charophytes and embryophytes (land plants) has been explained by a few chloroplast genomic characters including gene and intron (Manhart and Palmer 1990; Baldauf et al. 1990; Lew and Manhart 1993). Here we show that a charophyte, Spirogyra maxima, has the largest operon of angiosperm chloroplast genomes, rpl23 operon (trnⅠ-rpl23-rpl2-rps19-rpl22-rps3-rpl16-rpl14-rps8-infA-rpl36-rps11-rpoA) containing both embryophyte introns, rpl16.i and rpl2.i. The rpl23 gene cluster of Spirogyra contains a distinct eubacterial promoter sequence upstream of rpl23, which is the first gene of the green algal rpl23 gene cluster. This sequence is completely absent in angiosperms but is present in non-flowering plants. The results imply that, in the rpl23 gene cluster, early charophytes had at least two promoters, one upstream of trnⅠ and and another upstream of rpl23, which partially or completely lost its function in land plants. A comparison of gene clusters of prokaryotes, algal chloroplast DNAs and land plant cpDNAs indicated a loss of numerous genes in chlorophyll a+b eukaryotes. A phylogenetic analysis using presence/absence of genes and introns as characters produced trees with a strongly supported clade containing chlorophyll a+b eukaryotes. Spirogyra and embryophytes formed a clade characterized by the loss of rpl5 and rps9 and the gain of trnⅠ (CAU) and introns in rpl2 and rpl16. The analyses support the hypothesis that the rpl23 gene cluster and the rpl2 and rpl16 introns of land plants originated from a common ancestor of Spirogyra and land plants.

A Unique Gene Expression Signature of 5-fluorouracil

  • Kim, Ja-Eun;Yoo, Chang-Hyuk;Park, Dong-Yoon;Lee, Han-Yong;Yoon, Jeong-Ho;Kim, Se-Nyun
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.248-255
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    • 2005
  • To understand the response of cancer cells to anticancer drugs at the gene expression level, we examined the gene expression changes in response to five anticancer drugs, 5-fluorouracil, cytarabine, cisplatin, paclitaxel, and cytochalasin D in NCI-H460 human lung cancer cells. Of the five drugs, 5-fluorouracil had the most distinctive gene expression signature. By clustering genes whose expression changed significantly, we identified three clusters with unique gene expression patterns. The first cluster reflected the up-regulation of gene expression by cisplatin, and included genes involved in cell death and DNA repair. The second cluster pointed to a general reduction of gene expression by most of the anticancer drugs tested. A number of genes in this cluster are involved in signal transduction that is important for communication between cells and reception of extracellular signals. The last cluster represented reduced gene expression in response to 5-fluorouracil, the genes involved being implicated in DNA metabolism, the cell cycle, and RNA processing. Since the gene expression signature of 5-fluorouracil was unique, we investigated it in more detail. Significance analysis of microarray data (SAM) identified 808 genes whose expression was significantly altered by 5-fluorouracil. Among the up-regulated genes, those affecting apoptosis were the most noteworthy. The down-regulated genes were mainly associated with transcription-and translation-related processes which are known targets of 5-fluorouracil. These results suggest that the gene expression signature of an anticancer drug is closely related to its physiological action and the response of caner cells.