• Title/Summary/Keyword: rpoS

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Initial Characterization of yliH in Salmonella typhimurium

  • Park, Kyung-Hwa;Song, Mi-Ryung;Choy, Hyon-E.
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.558-565
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    • 2007
  • Using microarray analysis, we determined those Salmonella genes induced at the entry of stationary phase, and subsequently discovered that uncharacterized yliH was induced most dramatically. We set out to establish the molecular mechanism underlying the stationary phase induction of yliH under the standard culture condition, LB with vigorous aeration, by analyzing its promoter activity in various mutant backgrounds, lacking stationary phase ${\sigma}$, $RpoS^-$, or stringent signal molecules ppGpp, ${\Delta}relA$ ${\Delta}spoT$. It was found that the stationary phase induction of yliHp was partially dependent on rpoS but entirely dependent on ppGpp. DNA sequence analysis revealed that the Salmonella yliH gene is composed of 381 base-pair nucleotides, with overall amino acid sequence revealing 76.38% amino acid identity and 88.98% similarity with Escherichia coli yliH, although no motif from data base was noted for its possible role. Recently however, it has been reported that yliH in E. coli was implicated in biofilm formation and motility by repressing these activities (Domka et al., 2006). We have constructed a mutant Salmonella deleting yliH gene by allele replacement and examined its phenotype, and found that the yliH in Salmonella more or less affects motility and adherence by enhancing these activities. The effect on biofilm formation in Salmonella was uncertain. Moreover, addition of cloned yliH of E. coli into Salmonella did not reduce motility or adherence. Taken together, it appears that the pathways implicating yliH for biofilm formation and motility in E. coli and in Salmonella are somewhat different.

Effect of gacS and gacA Mutations on Colony Architecture, Surface Motility, Biofilm Formation and Chemical Toxicity in Pseudomonas sp. KL28

  • Choi, Kyung-Soon;Veeraragouda, Yaligara;Cho, Kyoung-Mi;Lee, Soo-O;Jo, Geuk-Rae;Cho, Kyung-Yun;Lee, Kyoung
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.492-498
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    • 2007
  • GacS and GacA proteins form a two component signal transduction system in bacteria. Here, Tn5 transposon gacS and gacA (Gac) mutants of Pseudomonas sp. KL28, an alkylphenol degrader, were isolated by selecting for smooth colonies of strain KL28. The mutants exhibited reduced ability to migrate on a solid surface. This surface motility does not require the action of flagella unlike the well-studied swarming motility of other Pseudomonas sp. The Gac mutants also showed reduced levels of biofilm and pellicle formation in liquid culture. In addition, compared to the wild type KL28 strain, these mutants were more resistant to high concentrations of m-cresol but were more sensitive to $H_2O_2$, which are characteristics that they share with an rpoS mutant. These results indicate that the Gac regulatory cascade in strain KL28 positively controls wrinkling morphology, biofilm formation, surface translocation and $H_2O_2$ resistance, which are important traits for its capacity to survive in particular niches.

Postsurgical Wound Infection Caused by Mycobacterium conceptionense Identified by Sequencing of 16S rRNA, hsp65, and rpoB Genes in an Immunocompetent Patient (16S rRNA, hsp65, 및 rpoB 염기순서분석으로 동정한 Mycobacterium conceptionense에 의한 면역능이 정상인 환자에서 발생한 수술후 창상감염)

  • Lee, Ja Young;Kim, Si Hyun;Shin, Jeong Hwan;Lee, Hyun-Kyung;Lee, Young Min;Song, Sae Am;Bae, Il Kwon;Kim, Chang-Ki;Jun, Kyung Ran;Kim, Hye Ran;Lee, Jeong Nyeo;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.17 no.1
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    • pp.23-27
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    • 2014
  • Rapidly growing mycobacteria are ubiquitous in the environment and are increasingly being recognized as opportunistic pathogens. Recently, a new species, Mycobacteium conceptionense, has been validated from the Mycobacterium fortuitum third biovariant complex by molecular analysis. However, there are few reports, and postsurgical wound infection by this species is rare. We report a case of postsurgical wound infection caused by M. conceptionense in an immunocompetent patient that was identified by a sequencing analysis of 16S rRNA, hps65, and rpoB genes.

Enterobacter cloacae, an Emerging Plant-Pathogenic Bacterium Affecting Chili Pepper Seedlings

  • Garcia-Gonzalez, Tanahiri;Saenz-Hidalgo, Hilda Karina;Silva-Rojas, Hilda Victoria;Morales-Nieto, Carlos;Vancheva, Taca;Koebnik, Ralf;Avila-Quezada, Graciela Dolores
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.1-10
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    • 2018
  • A previously unreported bacterial disease on chili pepper (Capsicum annuum L.) seedlings affecting as many as 4% of seedlings was observed in greenhouses in Chihuahua, Mexico (Delicias and Meoqui counties). Initial lesions appeared as irregular small spots on leaves and brown necrosis at margins tips were observed. Later, the spots became necrotic with a chlorotic halo. Advanced disease was associated with defoliation. A Gram negative, rod-shaped bacterium was isolated from diseased chili pepper seedlings. Three inoculation methods revealed that isolated strains produce foliage symptoms, similar to those observed in naturally infected seedlings. Pathogenic strains that caused symptoms in inoculated seedlings were re-isolated and identified to fulfill koch's postulate. Polyphasic approaches for identification including biochemical assays (API 20E and 50CH), carbon source utilization profiling (Biolog) and 16S rDNA, hsp60 and rpoB sequence analysis were done. Enterobacter cloacae was identified as the causal agent of this outbreak on chili pepper seedlings.

Morphological Characteristics of Needle Leaves and Analysis of Abies species based on Chloroplast DNA Sequences (한국 전나무(Abies holophylla), 일본 전나무(A. firma, A. homolepis), 그리고 법정 보호 전나무의 잎 형태적 특성 및 엽록체 DNA 분석)

  • Ahn, Chang Ho;Choi, Yong Eui;Park, Wan Geun;Han, Jung Yeon;Kwak, Yoo Shin;Kim, Se Chang;Park, Chan Woo
    • Journal of Korean Society of Forest Science
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    • v.108 no.2
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    • pp.200-207
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    • 2019
  • The aim of this study was to provide the basic information necessary to identify Korean fir (Abies holophylla), momi fir (A. firma), and Nikko fir (A. homolepis), and other fir trees planted in South Korea that are protected by law. Analysis of the morphological characteristics of the needles from each sample was investigated. The shape of the needle-leaf tip from A. holophylla was acute, whereas that from A. firma and A. homolepis was emarginate and that from the protected fir trees was obtuse. The number of stomata on the needles was not significantly different between A. holophylla and A. firma, and the number of stomata on the needles from A. homolepis and the protected fir trees were highly similar. In addition, the genetic differences among the Abies species were analyzed using the sequences of five chloroplast DNA regions-matK, atpF-atpH, rpoC2-rps2, rpoC1, and psbA-trnH.The atpF-atpH and psbA-trnH regions were useful for discriminating A. firma from the other species, but there were no differences among A. holophylla, A. homolepis, and the protected fir trees. The same chloroplast sequences were found in both A. holophylla and A. homolepis, which suggests that additional genetic studies might be necessary to identify the Abies species planted in both South Korea and Japan.

Comparative Transcriptome Analysis of Zerumbone-Treated Helicobacter pylori (Zerumbone 처리 헬리코박터 파이로리균의 전사체 분석 비교)

  • Woo, Hyun Jun;Yang, Ji Yeong;Kim, Sa-Hyun
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.301-309
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    • 2022
  • Helicobacter pylori (H. pylori) establishes infection in the human gastric mucosa for a long time and causes severe gastric diseases such as peptic ulcer and gastric cancer. When H. pylori is exposed to the antibacterial agents or inhibitors, the expression of pathogenic associated genes could be altered. In this study, we analyzed the transcriptional changes of H. pylori genes induced by zerumbone treatment. RNA expression changes were analyzed using next-generation sequencing (NGS), and then reverse transcription-polymerase chain reaction (RT-PCR) was performed to verify the results. As a result of NGS analysis, a total of 23 out of 1,632 genes were differentially expressed by zerumbone treatment. RT-PCR confirmed that zerumbone treatment regulated the expression level of 14 genes. Among the genes associated with DNA replication, transcription, virulence factors and T4SS components, 10 genes (dnaE, dnaQ, rpoA, rpoD, secA, flgE, flhA, virB5, virB8 and virB9) were significantly down-regulated and 4 genes (flaA, flaB, virB4 and virD4) were up-regulated. The results of our current study imply that zerumbone might be a potential therapeutic agent for H. pylori infection by regulating factors related to various H. pylori pathogenicity.

A two-component sensor kinase (GacS) mediated signal transduction pathway involved in production of antifungal compounds in Pseudomonas chlororaphis O6.

  • Kang, Beom-Ryong;Lee, Jung-Hoon;Kim, Hyun-Jung;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.99.1-100
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    • 2003
  • E. intermedium Blocontrol activity of a P. chlororaphis rhizobacteium O6, depends to the synthesis of extracellular secondary metabolites and exoenzymes, thought to antagonize the pathogenicity of a variety of phytopathogenic fungi. The production of secondary metabolites and exoenzymes in O6, depends essentially on the GacS-mediated signal transduction pathway, which activates largely unknown signal transduction pathway. To exploit the GacS-mediated signal transdcution pathway involved in activation of ph genes that are necessary for biosynthesis of phenazine from P. chlororaphis O6, we cloned and sequenced the phz operon, rpoS gene encoding stationary specific sigma factor, ppx gene encoding polyphosphatase, and lon gene encoding ion protease. Expression of each gene in wild type and GacS mutant were analyzed by RT-PCR. Transcripts from rpoS, phzI enconing acylhomoserine lactone (AHL) synthase, and ph structural genes in the GacS mutant were reduced in each of these growth phases compared to the wild type. The GacS or Lon mutant was found to be deficient in the production of phenzines, exoenzymes, and the acylhomoserine lactone. These mutants were not complemented by ph operon and addition of exogenous AHL. These results indicate that the GacS global regulatory systems controls phenazine production at multiple levels. Future research will focus to identifying the GacS-mediated regulatory cascade involving in production of phenazine in P. chlororaphis.

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Selection of Stable Reference Genes for Real-Time Quantitative PCR Analysis in Edwardsiella tarda

  • Sun, Zhongyang;Deng, Jia;Wu, Haizhen;Wang, Qiyao;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.112-121
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    • 2017
  • Edwardsiella tarda is a gram-negative pathogenic bacterium in aquaculture that can cause hemorrhagic septicemia in fish. Many secreted proteins have already been identified as virulent factors of E. tarda. Moreover, since virulent phenotypes are based on the expression regulation of virulent genes, understanding the expression profile of virulent genes is important. A quantitative RT-PCR is one of the preferred methods for determining different gene expressions. However, this requires the selection of a stable reference gene in E. tarda, which has not yet been systematically studied. Accordingly, this study evaluated nine candidate reference genes (recA, uup, rpoB, rho, topA, gyrA, groEL, rpoD, and 16S rRNA) using the Excel-based programs BestKeeper, GeNorm, and NormFinder under different culture conditions. The results showed that 16S rRNA was more stable than the other genes at different culture growth phases. However, at the same culture time, topA was identified as the reference gene under the conditions of different strains, different culture media, and infection, whereas gyrA was identified under the condition of different temperatures. Thus, in experiments, the expression of gapA and fbaA in E. tarda was analyzed by RT-qPCR using 16S rRNA, recA, and uup as the reference genes. The results showed that 16S rRNA was the most suitable reference gene in this analysis, and that using unsuitable reference genes resulted in inaccurate results.

The first case report on Vibrio atlanticus infection in cultured tiger puffer, Takifugu rubripes (우리나라 양식 자주복, Takifugu rubripes (Temminck & Schlegel, 1850)에서 발생한 Vibrio atlanticus의 첫 감염 사례 보고)

  • Lee, Nam-Sil;Cho, Miyoung;Jung, Sung-hee;Won, Kyoungmi
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.87-92
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    • 2018
  • A case of disease was occurred in tiger puffer, Takifugu rubripes, reared in circulated system (water temperature was about $20^{\circ}C$) on land in Korea. Diseased fish showed a unique external sign, the cloudy skin ulcer in various sizes. We investigated the cause of disease, and isolated one pure cultured bacterium, TP01. This had 99.7% homology with Vibrio atlanticus by sequence analysis of the 16S rRNA and rpoA genes. TP01 showed some differences with other reported V. atlanticus strains in the biochemical characteristics as like Voges-Proskauer test and gelatin hydrolysis.