• Title/Summary/Keyword: Virulence gene

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Raped Detection Method for Pathogenic Yersinia enterocolitica by a Multiplex-PCR (Multiplex-PCR을 이용한 병원성 Yersinia enterocolitica의 신속검출 기법에 관한 연구)

  • Lee, Young-Kee;Choi, Sung-Min;Oh, Soo-Kyung;Shin, Jei-Young;Ryeom, Kon
    • Journal of environmental and Sanitary engineering
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    • v.15 no.4
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    • pp.105-113
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    • 2000
  • We have isolated 65 strains(2.0%) of Y. enterocolitica among 3,219 water samples from 380 spring water sites in Seoul from 1994 to 1999. The biochemical characteristics of isolated strains revealed that TSI was A/A, urea, M.R.($37^{\circ}C$), nitrate, motility($37^{\circ}C$), sorbitol, maltose, manitol, arabinose, mannose, trehalose, xylose were positive(100%) and H$_2$S, arginine, lysine, oxidase, citrate, V.P.($37^{\circ}C$), DNase, motility($37^{\circ}C$), dulcitol, adonitol, lactose and raffinose were negative(100%). In in vitro virulence test, positive rate of AAG and CRMOX were 9.2% and 4.6%, respectively. However in the virulence gene detectable gene detectable test by multiplex-PCR using ail, yst, virF genes, 65 strains were all negative, meaning that Y.enterocolitica strains from domestic spring water were not detected for the virulence. Otherwise, mutiplex-PCR which using ail, yst and subgenus-specific primer pair was the best for identifying the virulence of Y. enterocolitica.

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Antimicrobial Resistance and Virulence Genes Presence in Escherichia coli Strains Isolated from Gomso Bay, Korea

  • Park, Kwon-Sam
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.221-227
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    • 2013
  • In total, 131 Escherichia coli isolates from surface seawater of the Gomso Bay, of Korea, were analyzed for their susceptibility to 22 different antimicrobials and for genes associated with antimicrobial resistance and virulence. According to the disk diffusion susceptibility test, the resistance to tetracycline was most prevalent (33.6%), followed by that to ampicillin (22.1%), ticarcillin (22.1%), and trimethoprim (16.8%). More than 46.6% of the isolates were resistant to at least one antimicrobial, and 22.9% were resistant to three or more classes of antimicrobials; these were consequently defined as multidrug resistant. We further found that 29 ampicillin-resistant isolates possessed genes encoding TEM-type (93.1%) and SHV-type (6.9%) ${\beta}$-lactamases. Among the 44 tetracycline-resistant isolates, tetA and tetC were found in 35 (79.5%) and 19 (43.2%), respectively, whereas tetB was detected in only three isolates (6.8%). With regard to virulence genes, merely 0.8% (n = 1) and 2.3% (n = 3) of the isolates were positive for the enteroaggregative E. coli-associated plasmid (pCVD432) gene and the enteropathogenic E. coli-specific attaching and effacing (eae) gene, respectively. Overall, these results not only provide novel insight into the necessity for seawater sanitation in Gomso Bay, but they help reduce the risk of contamination of antimicrobial-resistant bacteria.

Disruption of the metC Gene Affects Methionine Biosynthesis in Pectobacterium carotovorum subsp. carotovorum Pcc21 and Reduces Soft-Rot Disease

  • Seonmi, Yu;Jihee, Kang;Eui-Hwan, Chung;Yunho, Lee
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.62-74
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    • 2023
  • Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.

Disruption of the Dual Specificity Kinase Gene Causes the Reduction of Virulence in Candida albicans (이중특이성 인산화 효소의 결손이 Candida albicans 병원성에 미치는 효과)

  • Park, Yun-Hee;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.85-87
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    • 2011
  • The opportunistic human pathogen Candida albicans has the ability to convert from yeast-form to pseudohyphal or true hyphal form. The morphological transition is considered as an important virulence factor, because the decrease or lack in dimorphism causes the reduction of virulence. Our previous study revealed that the disruption of dual specificity kinase gene caused the reduction of dimorphism in C. albicans. Therefore we tested the effect of dual specificity kinase in virulence using mouse model. The mean survival time for kinase-defective strains was about 15 days in comparison with those of wild-type, 3.9 days. Moreover the fungal burden on kidneys for kinase-defective strains was decreased by ten-fold than that for wild-type. These results suggest possible involvement of dual specificity kinase in a novel signal transduction pathway for morphological transition and virulence of C. albicans.

Carbamoyl Phosphate Synthase Subunit CgCPS1 Is Necessary for Virulence and to Regulate Stress Tolerance in Colletotrichum gloeosporioides

  • Mushtaq, Aamar;Tariq, Muhammad;Ahmed, Maqsood;Zhou, Zongshan;Ali, Imran;Mahmood, Raja Tahir
    • The Plant Pathology Journal
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    • v.37 no.3
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    • pp.232-242
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    • 2021
  • Glomerella leaf spot (GLS) is a severe infectious disease of apple whose infective area is growing gradually and thus poses a huge economic threat to the world. Different species of Colletotrichum including Colletotrichum gloeosporioides are responsible for GLS. For efficient GLS control, it is important to understand the mechanism by which the cruciferous crops and C. gloeosporioides interact. Arginine is among one of the several types of amino acids, which plays crucial role in biochemical and physiological functions of fungi. The arginine biosynthesis pathway involved in virulence among plant pathogenic fungi is poorly understood. In this study, CgCPS1 gene encoding carbamoyl phosphate synthase involved in arginine biosynthesis has been identified and inactivated experimentally. To assess the effects of CgCPS1, we knocked out CgCPS1 in C. gloeosporioides and evaluated its effects on virulence and stress tolerance. The results showed that deletion of CgCPS1 resulted in loss of pathogenicity. The ∆cgcps1 mutants showed slow growth rate, defects in appressorium formation and failed to develop lesions on apple leaves and fruits leading to loss of virulence while complementation strain (CgCPS1-C) fully restored its pathogenicity. Furthermore, mutant strains showed extreme sensitivity to high osmotic stress displaying that CgCPS1 plays a vital role in stress response. These findings suggest that CgCPS1 is major factor that mediates pathogenicity in C. gloeosporioides by encoding carbamoyl phosphate that is involved in arginine biosynthesis and conferring virulence in C. gloeosporioides.

The Relationship of the Helicobacter pylori Virulence Factor Gene Subtype in Gastric Adenocarcinoma (위선암에서 Helicobacter pylori 독성인자와 유전자 아형의 관련성)

  • Shin Jong Min;Han Sang Young;Keum Dong Joo;Kim Kwang Jin;Jee Sam Ryong;Hong Gi Bong;Lee Jong Hun;Choi Seok Ryeol;Shin Woo Won
    • Journal of Gastric Cancer
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    • v.2 no.1
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    • pp.12-19
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    • 2002
  • Purpose: The H. pylori cagA gene, vacA gene and iceA gene are considered to be important virurence factors that have been implicated in the development of gastric adenocarcinoma. It was reported that the presence of IS605 elements may be responsible for rearrangements and lead to partial or total deletions of the cag pathogenicity island (PAI) and the virulence of cag PAI may be changed. However, different results regarding the association between these virulence factors and clinical disease have been reported from different geographic regions. This study evaluated the relationship between H. pylori virulence factors such as cagA, vacA, iceA, IS605 and gastric adenocarcinoma. Materials and Methods: H. pylori isolates were obtained from 54 infected patients (24 cases of gastric adenocarcinoma, 30 cases of control). H. pylori isolates were identified by PCR with ureC gene and 16S rRNA. PCR was performed to examine cagA, vacA, iceA and IS605 genotypes. Results: Significant difference was found in the negative rates of cagA between gastric adenocarcinoma group and control ($62.5\%\;vs.\;33.3\%$ P=0.033). No significant difference was found in the prevalence of iceA, vacA between gastric adenocar cinoma and control. The genotype of cagA+ vacA s1-m1 iceA1 was predominant in H. pylori isolates irrespective of the clinical outcome. IS605 in PAI was not found in gastric adenocarcinoma gruop and control. The positive rates of IS605 in genome were $33.3\%$ in gastric adenocarcinoma group and $36.7\%$ in control (P>0.05). In gastric carcinoma, the positive rate of $cagA^{+}/IS605$ was lower than in control ($12.5\%\;vs\;40.0\%$, P=0.025) and the positive rate of cagA-/IS605 was higher than in control ($54.2\%\;vs\;23.3\%$, P=0.02). Conclusion: H. pylori virulence factors had not related significantly with gastric adenocarcinoma. Further study is needed to examine the specificity of H. pylori strains.

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Effects of Agrobacterium tumefaciens and Tumor-inducing plasmid on the virulence gene expression (Agrobacterium tumefaciens와 Tumor-inducing 플라스미드에 의한 virulence 유전자의 발현)

  • Eum, Jin-Seong
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.4
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    • pp.1000-1006
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    • 2011
  • We examined the effects of various phenolic compounds, Ti plasmids(octopine, nopaline) and A. tumefaciens on the vir gene expression. Nine phenolic compounds were tested using 3 types of Ti plasmid and 3 strains of A. tumefaciens on the vir gene expression. It was also investigated how the levels of vir gene expression could be related to specific phenolic compounds. Six phenolic compounds as 4-hydroxyacetophenone, phenol, catechol, resorcinol, acetosyringone and vanillin had exhibited a strong effect on the vir gene expression of A. tumefaciens MW107 containing nopaline Ti plasmid. The vir genes of A. tumefaciens MW105 and MW108 containing octopine Ti plasmids were remarkably stimulated only by acetosyringone. Thus, it appeared that the vir gene inducing abilities were differed by the kinds of phenolic compounds, A. tumefaciens and Ti plasmids.

Enhanced Homologous Recombination in Fusarium verticillioides by Disruption of FvKU70, a Gene Required for a Non-homologous End Joining Mechanism

  • Choi, Yoon-E.;Shim, Won-Bo
    • The Plant Pathology Journal
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    • v.24 no.1
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    • pp.1-7
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    • 2008
  • Fusarium verticillioides (teleomorph Gibberella moniliformis) is associated with maize worldwide causing ear rot and stalk rot, and produces fumonisins, a group of mycotoxins detrimental to humans and animals. While research tools are available, our understanding of the molecular mechanisms associated with fungal virulence and fumonisin biosynthesis in F. verticillioides is still limited. One of the restraints that hampers F. verticillioides gene characterization is the fact that homologous recombination (HR) frequency is very low (<2%). Screening for a true gene knock-out mutant is a laborious process due to a high number of ectopic integrations. In this study, we generated a F. verticillioides mutant (SF41) deleted for FvKU70, a gene directly responsible for non-homologous end-joining mechanism, with the aim of improving HR frequency. Here, we demonstrate that FvKU70 deletion does not impact key Fverticillioides phenotypes, e.g., development, secondary metabolism, and virulence, while dramatically improving HR frequency. Significantly, we also confirmed that a high percentage (>85%) of the HR mutant strains harbor a desired mutation with no additional copy of the mutant allele inserted in the genome. We conclude that SF41 is suitable for use as a type strain when performing high-throughput gene function studies in F. verticillioides.

Effect of Sonication and vir Genes on Transient Gene Expression in Agrobacterium-Mediated Transformation (Agrobacterium을 이용한 형질전환에서 sonication과 vir 유전자들의 효과)

  • 이병무
    • Journal of Life Science
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    • v.11 no.4
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    • pp.316-320
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    • 2001
  • Sonication tremendously improves the efficiency of Agrobacterium infection by introducing small and uniform fissures and channels throughout the targeted tissue. Using shoot tips of cotton as explants, the effect of sonication treatment and virulence genes in Agrobacterium tumefaciens on transformation efficiency was investigated. The pat gene which encodes resistance to the herbicide, glufosinate, was used as a selectable marker. Transformation efficiency was evaluated on th basis of survival rates of cocultivated shoot tips on selection medium containing 2.5 mg/l gulfosinate-ammonium(ppt) adn 25. mg/l Clavamax. Sonication from 5 to 15 second has a positive effect on shoop tip survival. However, whil virE as well as virG or vir GN54D showed an enhancement in transformation efficiency, virE,. virG resulted in the most significant enhancement. Overall, the combination of additional virG/virE gene and sonication treatment resulted in the most significant increase in transformation efficiency.

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DD1.5k, the Gene Preferentially Expressed in Bloodstream Isolates of Vancomycin-Resistant Enterococcus faecium

  • Kim, Seung-Han;Lee, Dong-Gun;Yoo, Jin-Hong;Park, Su-Mi;Park, Jung-Hyun;Shin, Wan-Shik;Lee, Kyungwon;Dongeun Yong;Lee, Wee-Gyo
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.143-146
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    • 2004
  • Vancomycin-resistant Enterococcus faecium (VREFM) is becoming a threatening pathogen. We identified a gene called DD1.5K by differential display-PCR, which was preferentially expressed in the bloodstream isolates of VREFM. Due to its amino acid similarity to transfer complex protein, trsE, and tissue-specific expression, this gene may be involved in virulence of VREFM.