• Title/Summary/Keyword: biological resistance analysis

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Subcellular Localization of Catalase Encoded by the ctl+ Gene in Schizosaccharomyces pombe

  • Lee, Sang-il;Lee, Joon;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.156-159
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    • 2000
  • The cttl+ gene in Schizosaccharomyces pombe encoeds a catalse responsible for H2O2-resistance of this organism as judged by the H2O2-sensitive phenotype of the ctt1Δ mutant. In this study, we investigated the subcellular localization of the Ctt1 gene product. In wild type cells catalase activity was detected in the organelle fraction as well as in the cytosol. The ctt1Δ mutant contained no catalase activity, indicating that both cytosolic and organellar catalases are the products of a single ctt1+ gene. Western bolt analysis revealed two catalase bands, both of which disappeared in the ctt1Δ mutant. The major, fastermigrating band existed in the cytosol whereas the monor, slower-migrating band appeared to be located in organelles, most likely in peroxisomes. These results suggest that the ctt1+ gene product targeted to the peroxisome is a modified form of the one in the cytosol.

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Community Structure and Cluster Analysis of the Benthic Macroinvertebrates in Inflow and Outflow area of Ten Reservoirs of the Nakdong River System (낙동강수계 10개 호소의 유입 및 유출부의 저서성대형무척추동물 군집구조 및 유사도 분석)

  • Lee, Mi-Jin;Park, Jin-Young;Seo, Jung-Kwan;Lee, Hae-Jin;Seo, Eul-Won;Lee, Jong-Eun
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1758-1763
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    • 2009
  • Biological assessments of benthic macroinvertebrates community structure were carried out in ten reservoir inflow and outflow areas of the Nakdong river system from April 2008 to August 2008. The collected benthic macroinvertebrates from the surveyed sites were 9,427.6 inds./$m^2$, 42 species belonging to 29 families and 16 orders. A dominant species was Chironomus sp. and a subdominant species was Ecdyonurus levis. In the all sites, Ephemeroptera had the most number of species (17.81%) and Diptera had the most number of individuals (64.00%). Dominance, species diversity, species richness, and species evenness indexes were 0.68, 2.42, 2.96 and 0.54, respectively. The numbers of benthic macroinvertebrates species in each surveyed site were from 21 (site 9 Jilnal wetland) to 51 (site 2 Yeongcheon reservoir). As a result of an analysis about relative resistance and resilience of stability factors, the most popular group was group I with high indices in both resistance and resilience. Site 7 (Beongae wetland) and site 8 (Jangcheok reservoir) had the highest similarity in the result of cluster analysis using the surveyed benthic macroinvertebrates.

Genetic Analysis and Characterization of a Bacteriophage ØCJ19 Active against Enterotoxigenic Escherichia coli

  • Kim, Gyeong-Hwuii;Kim, Jae-Won;Kim, Jaegon;Chae, Jong Pyo;Lee, Jin-Sun;Yoon, Sung-Sik
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.746-757
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    • 2020
  • Enterotoxigenic Escherichia coli (ETEC) is the major pathogenic E. coli that causes diarrhea and edema in post-weaning piglets. In this study, we describe the morphology and characteristics of ØCJ19, a bacteriophage that infects ETEC, and performed genetic analysis. Phage ØCJ19 belongs to the family Myoviridae. One-step growth curve showed a latent phase of 5 min and burst size of approximately 20 phage particles/infected cell. Phage infectivity was stable for 2 h between 4℃ and 55℃, and the phage was stable between pH 3 and 11. Genetic analysis revealed that phage ØCJ19 has a total of 49,567 bases and 79 open reading frames (ORFs). The full genomic sequence of phage ØCJ19 showed the most similarity to an Escherichia phage, vB_EcoS_ESCO41. There were no genes encoding lysogeny, toxins, virulence factors, or antibiotic resistance in this phage, suggesting that this phage can be used safely as a biological agent to control ETEC. Comparative genomic analysis in terms of the tail fiber proteins could provide genetic insight into host recognition and the relationship with other coliphages. These results showed the possibility to improve food safety by applying phage ØCJ19 to foods of animal origin contaminated with ETEC and suggests that it could be the basis for establishing a safety management system in the animal husbandry.

Mutation in the rpoB Gene of Mycobacterium leprae from Korean Laprosy Patients

  • Kim, Soon-Ok;chae, Gue-Tae;Shin, Hang-Kye;Kim, Nan-Hee;Lee, In-Hyung;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.287-293
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    • 2001
  • A fast and easy PCR-SSCP method was developed and assessed for the early detection of rifampin-resistant Mycobacterium leprae in skin biopsy samples from Korean leprosy patients. The 190 bp of the rpoB gene, in which mutation is known to cause resistance to rifampin, was amplified by PCR and then analyzed by SSCP and DNA sequencing, All PCR products showing mobility shift on PCR-SSCP contained mutations, demonstrating that this method can be used for an early diagnositic method to detect a putative rifampin-resistant M. leprae strain. DNA sequence analysis revealed that 19 of 34 patient samples contained M. leprae strains with missense mutations in the rpoB gene: five were the same mutations previously reported to cause rifampin resistance and eight were the new type of mutatios that likely cause rifampin resistance. These newly identified dmutations, whose all five cytosine bases of four amino acids were substitued with thymine, were found at different sites from those reported in Mycobacterium tuberculosis or M. leprae. Therefore, they may provide additional clues to understand the molecular biological basis on the rifampin resistance of M. leprae.

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L-glutamine:D-fructose-6-phosphate Aminotransferase as a Key Protein Linked to Multidrug Resistance in E. coli KD43162

  • Lee, Sung-Eun;Jung, Tae-Jeon;Park, Byeoung-Soo;Kim, Byung-Woo;Lee, Eun-Woo;Kim, Hye Jin;Yum, Jong Hwa
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.227-232
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    • 2015
  • A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

Isolation and Characterization of the Biosynthetic Gene Clusters for Aminoglycoside Antibiotics

  • Jung Yong-Gyun;Jo You-Young;Hyun Chang-Gu;Lee In Hyung;Yang Young-Ye1l;Suh Joo-Won
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.146-156
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    • 2001
  • The biosynthetic gene clusters for bluensomycin and spectinomycin were isolated and characterized from the bluensomycin producer, Streptomyces bluensis ATCC27420 and the spectinomycin producer, Streptomyces spectabilis ATCC27741, respectively. PCR primers were designed specifically to amplify a segment of dTDP-glucose synthase gene based on its conserved sequences of several actinomycete strains. By screening cosmid libraries using amplified PCR fragments, 30-kb and 45-kb DNA fragments were isolated from Streptomyces bluensis and Streptomyces spectabilis, respectively. Sequencing analysis of them revealed that each contains 15 open reading frames (ORFs). Some of these ORFs were turned out to be antibiotic resistance genes (blmA and speN), dTDP-glucose synthase genes (blmD and spcD), and dTDP-D-glucose 4,6-dehydratase genes (blmE and spcE), suggesting that the blm and spec gene clusters are likely involved in the biosynthesis of bluensomycin and spectinomycin, respectively.

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Message in a Bottle: Chemical Biology of Induced Disease Resistance in Plants

  • Schreiber, Karl;Desveaux, Darrell
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.245-268
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    • 2008
  • The outcome of plant-pathogen interactions is influenced significantly by endogenous small molecules that coordinate plant defence responses. There is currently tremendous scientific and commercial interest in identifying chemicals whose exogenous application activates plant defences and affords protection from pathogen infection. In this review, we provide a survey of compounds known to induce disease resistance in plants, with particular emphasis on how each compound was originally identified, its putative or demonstrated mechanism of defence induction, and the known biological target(s) of each chemical. Larger polymeric structures and peptides/proteins are also discussed in this context. The quest for novel defence-inducing molecules would be aided by the capability for high-throughput analysis of candidate compounds, and we describe some issues associated with the development of these types of screens. Subsequent characterization of hits can be a formidable challenge, especially in terms of identifying chemical targets in plant cells. A variety of powerful molecular tools are available for this characterization, not only to provide insight into methods of plant defence activation, but also to probe fundamental biological processes. Furthermore, these investigations can reveal molecules with significant commercial potential as crop protectants, although a number of factors must be considered for this potential to be realized. By highlighting recent progress in the application of chemical biology techniques for the modulation of plant-pathogen interactions, we provide some perspective on the exciting opportunities for future progress in this field of research.

Genome-Based Insights into the Thermotolerant Adaptations of Neobacillus endophyticus BRMEA1T

  • Lingmin Jiang;Ho Le Han;Yuxin Peng;Doeun Jeon;Donghyun Cho;Cha Young Kim;Jiyoung Lee
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.321-329
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    • 2023
  • The bacterium Neobacillus endophyticus BRMEA1T, isolated from the medicinal plant Selaginella involvens, known as its thermotolerant can grow at 50℃. To explore the genetic basis for its heat tolerance response and its potential for producing valuable natural compounds, the genomes of two thermotolerant and four mesophilic strains in the genus Neobacillus were analyzed using a bioinformatic software platform. The whole genome was annotated using RAST SEED and OrthVenn2, with a focus on identifying potential heattolerance-related genes. N. endophyticus BRMEA1T was found to possess more stress response genes compared to other mesophilic members of the genus, and it was the only strain that had genes for the synthesis of osmoregulated periplasmic glucans. This study sheds light on the potential value of N. endophyticus BRMEA1T, as it reveals the mechanism of heat resistance and the application of secondary metabolites produced by this bacterium through whole-genome sequencing and comparative analysis.

SETDB1 mediated FosB expression increases the cell proliferation rate during anticancer drug therapy

  • Na, Han-Heom;Noh, Hee-Jung;Cheong, Hyang-Min;Kang, Yoonsung;Kim, Keun-Cheol
    • BMB Reports
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    • v.49 no.4
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    • pp.238-243
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    • 2016
  • The efficacy of anticancer drugs depends on a variety of signaling pathways, which can be positively or negatively regulated. In this study, we show that SETDB1 HMTase is down-regulated at the transcriptional level by several anticancer drugs, due to its inherent instability. Using RNA sequence analysis, we identified FosB as being regulated by SETDB1 during anticancer drug therapy. FosB expression was increased by treatment with doxorubicin, taxol and siSETDB1. Moreover, FosB was associated with an increased rate of proliferation. Combinatory transfection of siFosB and siSETDB1 was slightly increased compared to transfection of siFosB. Furthermore, FosB was regulated by multiple kinase pathways. ChIP analysis showed that SETDB1 and H3K9me3 interact with a specific region of the FosB promoter. These results suggest that SETDB1-mediated FosB expression is a common molecular phenomenon, and might be a novel pathway responsible for the increase in cell proliferation that frequently occurs during anticancer drug therapy.

Isolation and characterization of induced disease resistance (ISR)-deficient mutants of a biocontrol bacterium Pseudomonas chlororaphis O6.

  • Han, Song-Hee;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.101.1-101
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    • 2003
  • Lipopolysaccharide, siderophore, and cyclic dipeptide have been shown to be necessary for ISR induction by pseudomnads. However, there is no report on cloning of genes or generating specific mutants involving in ISR activity. A biological control bacteium P. chlororaphis O6 induces resistance to Erwinia carotovora subsp. carotovara SCCI in tobacco and induces drought resistance in Arabidopsis. To isolate genes involved in ISR activity and induction of drough resistance of O6, we constructed Tn5 mutants and were used to screen for ISR activity and drought resistance activity using microtiter assay with tobacco and Arabidopsis. Thirty-three ISR-deficient mutants were selected, and the nine ISR-deficient mutants were also lost activity of drought resistance. The flanking sequence analysis of the ISR and drought resistance-deficient mutants showed that a gacS gene encoding a two-component sensor kinase, and a mce gene encoding a protein involved in mycobacterial cell entry were mutated. The flanking sequence of each Tn5 mutant altered ISR activity is currently under investigation. These results indicate that gacS and mce are important genes in induction of ISR activity and drought resistance of P. chlororaphis O6. Our works will open opportunities for identification of bacterial genes or traits that are involved in ISR activity and induced drought resistance of P. chlororaphis O6.

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