• Title/Summary/Keyword: R-gene

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Characterization of Cytophaga-Flavobacteria Community Structure in the Bering Sea by Cluster-specific 16S rRNA Gene Amplification Analysis

  • Chen, Xihan;Zeng, Yonghui;Jiao, Nianzhi
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.194-198
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    • 2008
  • A newly designed Cytophaga-Flavobacteria-specific 16S rRNA gene primer pair was employed to investigate the CF community structure in the Bering Sea, revealing a previously unknown and unexpected high CF diversity in this high latitude cold sea. In total, 56 clones were sequenced and 50 unique CF 16 rRNA gene fragments were obtained, clustering into 16 CF subgroups, including nine cosmopolitan subgroups, five psychrophilic subgroups, and two putatively autochthonous subgroups. The majority of sequences (82%) were closely related to uncultured CF species and could not be classified into known CF genera, indicating the presence of a large number of so-far uncultivated CF species in the Bering Sea.

Genetic Identification of the Kimchi Strain Using PCR-based PepN and 16S rRNA Gene Sequence (PepN과 16S rRNA Gene Sequence 및 PCR 방법을 이용한 김치 젖산균의 동정)

  • Lee, Myung-Ki;Park, Wan-Soo;Lee, Byong-H.
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1331-1335
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    • 2000
  • The WL6 strain isolated from Kimchi could not be made scientific name because it was identified as three species, i.e., Leuconostoc mesenternides ssp cremoris, Leu. mesenteroides ssp. dextranicum or Lactobacillus bifermentans when it was tested by API kit or Biolog system methods. The unidentifiable WL6 strain was finally reclassified as Lactobacillus bifermentans by genetic identification using two PCR-based specific sequence primer sets which were originated from homologous pepN and 16S rRNA genes.

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Uridylate kinase as a New Phylogenetic Molecule for Procaryotes

  • Lee, Dong-Geun;Lee, Jin-Ok;Lee, Jae-Hwa
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.810-814
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    • 2003
  • For the phylogenetic analysis of procaryotes, 16S rRNA gene has been used. In spite of it's common use, so high conservative of 16S rRNA gene limited resolving power, hence other molecule was applied in this study and the result was compared with that of 16S rRNA. COG (Clusters of Orthologous of protein) algorithm revealed that three COGs were only detected in 42 procaryotes ; transcription elongation factor (COG0195), bacterial DNA primase (COG0358) and uridylate kinase (COG0528). Uridylate kinase gene was selected owing to the similarity and one single copy number in each genome. Phylogenetic tree of 16S rRNA gene and uridylate kinase showed similarities and differences. Uridylate kinase may help the problem of very high conservative of 16S rRNA gene in rhylogenetic analysis and it would help to access more accurate discrimination and phylogenetic analysis of bacteria.

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Enhancer Function of MicroRNA-3681 Derived from Long Terminal Repeats Represses the Activity of Variable Number Tandem Repeats in the 3' UTR of SHISA7

  • Lee, Hee-Eun;Park, Sang-Je;Huh, Jae-Won;Imai, Hiroo;Kim, Heui-Soo
    • Molecules and Cells
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    • v.43 no.7
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    • pp.607-618
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    • 2020
  • microRNAs (miRNAs) are non-coding RNA molecules involved in the regulation of gene expression. miRNAs inhibit gene expression by binding to the 3' untranslated region (UTR) of their target gene. miRNAs can originate from transposable elements (TEs), which comprise approximately half of the eukaryotic genome and one type of TE, called the long terminal repeat (LTR) is found in class of retrotransposons. Amongst the miRNAs derived from LTR, hsa-miR-3681 was chosen and analyzed using bioinformatics tools and experimental analysis. Studies on hsa-miR-3681 have been scarce and this study provides the relative expression analysis of hsa-miR-3681-5p from humans, chimpanzees, crab-eating monkeys, and mice. Luciferase assay for hsa-miR-3681-5p and its target gene SHISA7 supports our hypothesis that the number of miRNA binding sites affects target gene expression. Especially, the variable number tandem repeat (VNTR) and hsa-miR-3681-5p share the binding sites in the 3' UTR of SHISA7, which leads the enhancer function of hsamiR-3681-5p to inhibit the activity of VNTR. In conclusion, hsa-miR-3681-5p acts as a super-enhancer and the enhancer function of hsa-miR-3681-5p acts as a repressor of VNTR activity in the 3' UTR of SHISA7.

Expression of B Cell Activating Factor Pathway Genes in Mouse Mammary Gland

  • Choi, S.;Jung, D.J.;Bong, J.J.;Baik, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.153-159
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    • 2007
  • In our previous study, overexpression of extracellular proteinase inhibitor (Expi) gene accelerated apoptosis of mammary epithelial cells, and induced expression of B cell activating factor (BAFF) gene. In this study, we found induction of BAFF-receptor (BAFF-R) gene expression in the Expi-transfected cells. A proliferation-inducing ligand (APRIL) gene is another TNF family member and the closest known relative of BAFF. We found induction of APRIL gene expression in the Expi-overexpressed apoptotic cells. NF-${\kappa}$B gene was also induced in the Expi-overexpressed cells. Expression patterns of BAFF and APRIL pathway-related genes were examined in in vivo mouse mammary gland at various reproductive stages. Expression levels of BAFF gene were very low at early pregnancy, increased from mid-pregnancy, and peaked at lactation, and thereafter decreased at involution stages of mammary gland. Expression of BAFF-R gene was highly induced in involution stages compared to lactation stages. Thus, expression patterns of BAFF-R gene were correlated to apoptotic status of mammary gland: active apoptosis of mammary epithelial cells occurs at involution stage of mammary gland. Expression levels of NF-${\kappa}$B gene were higher in involution stages compared to lactation stages. We analyzed mRNA levels of bcl-2 family genes from different stages of mammary development. Bcl-2 gene expression was relatively constant during lactation and involution stages. There was a slight increase in bcl-xL gene expression in involution stages compared to lactation state. Bax gene expression was highly induced in involution stage. Our results suggest that signaling pathways activated by both BAFF and ARRIL in mammary gland point towards NF-${\kappa}$B activation which causes upregulation of bax.

Sequence Analysis and Functional Expression of the Structural and ]Regulatory Genes for Pyruvate Dehydrogenase

  • Hwan Youn;Jangyul Kwak
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.43-50
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    • 2002
  • A cluster of genes encoding the pyruvate dehydrogenase complex (PDC) of Streptomyces seoulensis, a Gram-positive bacterium, was cloned and sequenced. The genes of S. seoulensis consist of four open reading frames. The first gene, lpd, which encodes a lipoamide dehydrogenase, is followed by pdhB encoding a dihydrolipoamide acetyltransferase (E2p), pdhR, a regulatory gene, and pdhA encoding a pyruvate dehydrogenase component (Elp). Elp had an unusual homodimeric subunit, which has been known only in Gram-negative bacteria S. seoulensis E2p contains two lipoyl domains like those of humans and Streptomyces faecalis. The pdhR gene appears to be clustered with the structural genes of S. seoulensis PDC. The PdhR-overexpressed S. seoulensis howed growth retardation and the decrease of Elp, indicating that PdhR regulates the function of PDC by repressing the expression of Elp. A strain of Streptomyces licidans overexpressing S. seoulensis PdhR showed a significant decreasein the level of actinorhodin, implying a regulatory role for Streptomyces PDC in antibiotic biosynthesis.

Gene Mutations of 23S rRNA Associated with Clarithromycin Resistance in Helicobacter pylori Strains Isolated from Korean Patients

  • Kim, Jung-Mogg;Kim, Joo-Sung;Kim, Na-Young;Kim, Yeoung-Jeon;Kim, In-Young;Chee, Young-Joon;Lee, Chul-Hoon;Jung, Hyun-Chae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1584-1589
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    • 2008
  • Although resistance of Helicobacter pylori to clarithromycin is a major cause of failure of eradication therapies, little information is available regarding gene mutations of clarithromycin-resistant primary and secondary H. pylori isolates in Korea. In the present study, we examined gene mutations of H. pylori 238 rRNA responsible for resistance to clarithromycin. DNA sequences of the 238 rRNA gene in 21 primary clarithromycin-resistant and 64 secondary clarithromycin-resistant strains were determined by PCR amplification and nucleotide sequence analyses. Two mutations of the 238 rRNA gene, A2143G and T2182C, were observed in primary clarithromycin-resistant isolates. In secondary isolates, dual mutation of A2143G+T2182C was frequently observed. In addition, A2143G+T2182C+ T2190C, A2143G+T2182C+C2195T, and A2143G+T2182C+A2223G were observed in secondary isolates. Furthermore, macrolide binding was tested on purified ribosomes isolated from T2182C or A2143C mutant strains with $[^{14}C]$erythromycin. Erythromycin binding increased in a dose-dependent manner for the susceptible strain but not for the mutant strains. These results indicate that secondary isolates show a greater variety of 238 rRNA gene mutation types than primary isolates, and triple mutations of secondary isolates are associated with A2143G+T2182C in H. pylori isolated from Korean patients.

A Mutation of a Putative NDP-Sugar Epimerase Gene in Ralstonia pseudosolanacearum Attenuates Exopolysaccharide Production and Bacterial Virulence in Tomato Plant

  • Hyoung Ju Lee;Sang-Moo Lee;Minseo Choi;Joo Hwan Kwon;Seon-Woo Lee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.417-429
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    • 2023
  • Ralstonia solanacearum species complex (RSSC) is a soil borne plant pathogen causing bacterial wilt on various important crops, including Solanaceae plants. The bacterial pathogens within the RSSC produce exopolysaccharide (EPS), a highly complicated nitrogencontaining heteropolymeric polysaccharide, as a major virulence factor. However, the biosynthetic pathway of the EPS in the RSSC has not been fully characterized. To identify genes in EPS production beyond the EPS biosynthetic gene operon, we selected the EPS-defective mutants of R. pseudosolanacearum strain SL341 from Tn5-inserted mutant pool. Among several EPSdefective mutants, we identified a mutant, SL341P4, with a Tn5-insertion in a gene encoding a putative NDP-sugar epimerase, a putative membrane protein with sugar-modifying moiety, in a reverse orientation to EPS biosynthesis gene cluster. This protein showed similar to other NDP-sugar epimerases involved in EPS biosynthesis in many phytopathogens. Mutation of the NDP-sugar epimerase gene reduced EPS production and biofilm formation in R. pseudosolanacearum. Additionally, the SL341P4 mutant exhibited reduced disease severity and incidence of bacterial wilt in tomato plants compared to the wild-type SL341 without alteration of bacterial multiplication. These results indicate that the NDP-sugar epimerase gene is required for EPS production and bacterial virulence in R. pseudosolanacearum.

A Report of Eighteen Unrecorded Fungal Species in Korea (국내에서 분리된 미기록 진균 18종 보고)

  • Ahn, Geum Ran;Choi, Min Ah;Kim, Ji Eun;Seo, Eun Ji;Kim, Jun Young;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.292-303
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    • 2017
  • During a survey of culturable fungi in Korea, 18 unrecorded fungal species were isolated and identified from the indoor air of mushroom cultivation houses, the materials used for preparation of mushroom cultivation media, wild plants, and funitures. This study reports the descriptions of the 18 unrecorded fungal species: Aspergillus creber, Ceratocystis paradoxa, Colletotrichum spaethianum, Coniochaeta velutina, Coprinellus xanthothrix, Epicoccum sorghinum, Leptosphaeria rubefaciens, Myrothecium gramineum, Paraconiothyrium fuckelii, Penicillium erubescens, Penicillium melinii, Penicillium pulvillorum, Penicillium sabulosum, Penicillium turbatum, Pestalotiopsis portugalica, Pilidiella castaneicola, Rachicladosporium pini, and Umbelopsis nana. For all the identified species, the morphological characteristics including the features of colony formed on media, images of light microscopy, and molecular phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer ribosomal DNA (ITS rDNA), 18S rDNA, 28S rDNA, ${\beta}-tubulin$ gene, calmodulin gene, and translation elongation factor gene were described.

Identification of MC1R gene variants of Hanwoo and Holstein meat using PCR-RFLP (PCR-RFLP를 이용한 한우와 젖소고기의 MC1R 유전자변이 검출)

  • Koh Ba-Ra-Da;Kim Yong-Hwan;Park Seong-Do;Na Ho-Myung;Kim Jeong-Nam;Sung Chang-Min;Lee Sam-Soo
    • Korean Journal of Veterinary Service
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    • v.28 no.3
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    • pp.259-265
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    • 2005
  • The melanocortin 1 receptor (MC1R) encoded by the coat color extension gene (E) plays a key role in the signaling pathway of melanin synthesis. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no. Y19103). A size of 483bp (482bp for Hanwoo) was amplified by PCR, digested with Hpa II restriction enzyme and electrophoresed in $1.5\%$ agarose gel. When the amplified DNA product (483 bp) was digested with Hpa II restriction enzyme, Hanwoo meat showed a single band of 482bp, whereas two fragments of 325bp and 158bp were detected in Holstein, Angus and meat of Hanwoo / Holstein cross cow having back coat color phenotype, respectively. The results of this experiment Indicate that new designed primers of bovine MCIR gene may be useful for identification of Hanwoo meat from Holstein, Black Angus and Hanwoo / Holstein cross cow meat.