• Title/Summary/Keyword: Bacterial DNA

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Imidazole Ring-Opened DNA Purines and Their Biological Significance

  • Barbara, Tudek
    • BMB Reports
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    • v.36 no.1
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    • pp.12-19
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    • 2003
  • Fragmentation of purine imidazole ring and production of formamidopyrimidines in deoxynucleosides (Fapy lesions) occurs upon DNA oxidation as well as upon spontaneous or alkali-triggered rearrangement of certain alkylated bases. Many chemotherapeutic agents such as cyclophosphamide or thiotepa produce such lesions in DNA. Unsubstituted FapyA and FapyG, formed upon DNA oxidation cause moderate inhibition of DNA synthesis, which is DNA polymerase and sequence dependent. Fapy-7MeG, a methylated counterpart of FapyG-, a efficiently inhibits DNA replication in vitro and in E.coli, however its mutagenic potency is low. This is probably due to preferential incorporation of cytosine opposite Fapy-7MeG and preferential extension of Fapy-7MeG:C pair. In contrast, FapyA and Fapy-7MeA possess miscoding potential. Both lesions in SOS induced E.coli preferentially mispair with cytosine giving rise to A$\rightarrow$G transitions. Fapy lesions substituted with longer chain alkyl groups also show simult aneous lethal and mutagenic properties. Fapy lesions are actively eliminated from DNA by repair glycosylases specific for oxidized purines and pyrimidines both in bacteria and eukaryotic cells. Bacterial enzymes include E.coli formamidopyrimidine-DNA-glycosylase (Fpg protein), endonuclease III (Nth protein) and endonuclease VIII (Nei protein).

Detection of Aromatic Pollutants by Bacterial Biosensors Bearing Gene Fusions Constructed with the dnaK Promoter of Pseudomonas sp. DJ-12

  • Park, Sang-Ho;Lee, Dong-Hun;Oh, Kye-Heon;Lee, Kyoung;Kim, Chi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.417-422
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    • 2002
  • Gene fusions were constructed by the transcriptional fusion of the dnaK promoter of pseudomonas sp. DJ-12 or E. coli to the lux or luc marker gene. The dnaKp-DJ::luxCDABE bioluminescent fusion in the biosensor using the Pseudomonas sp. DJ-12 dnaK promoter exhibited about 5-fold more extensive response to ethanol than that of dnaKp-EC::luxCDABE. The bioluminescent response of the dnaK-DJ::luc fusion to ethanol was much weaker than those of the other fusions. The biosensor harboring the dnaKp-DJ::luCDABE fusion was examined for its bioluminescence production based on exposure to aromatic compounds, such as biphenyl, 4-chlorobiphenyl (4CB), 4-hydroxybenzoate (4HBA), and catechol. In particular, the bioluminescence produced by the dnaKp-DJ::luxCDABE fusion was most sensitive to 1 mM biphenyl and 4CB when exposed for 80 min, and the responses were also very strong to other aromatics. Therefore, the biosensor bearing the dnaKp-DJ::luxCDABE fusion would appear to be the most useful for the detection of aromatics and other pollutants.

PCR-based Specific Detection of Ralstonia solanacearum by Amplification of Cytochrome c1 Signal Peptide Sequences

  • Kang, Man-Jung;Lee, Mi-Hee;Shim, Jae-Kyung;Seo, Sang-Tae;Shrestha, Rosemary;Cho, Min-Seok;Hahn, Jang-Ho;Park, Dong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1765-1771
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    • 2007
  • A polymerase chain reaction (PCR)-based method was developed to detect the DNA of Ralstonia solanacearum, the causal agent of bacterial wilt in various crop plants. One pair of primers (RALSF and RALSR), designed using cytochrome c1 signal peptide sequences specific to R. solanacearum, produced a PCR product of 932 bp from 13 isolates of R. solanacearum from several countries. The primer specificity was then tested using DNA from 21 isolates of Ralstonia, Pseudomonas, Burkholderia, Xanthomonas, and Fusarium oxysporum f. sp. dianthi. The specificity of the cytochrome c1 signal peptide sequences in R. solanacearum was further confirmed by a DNA-dot blot analysis. Moreover, the primer pair was able to detect the pathogen in artificially inoculated soil and tomato plants. Therefore, the present results indicate that the primer pair can be effectively used for the detection of R. solanacearum in soil and host plants.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems

  • Choi, Hae-Yeon;Kim, Youn-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.285-292
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    • 2008
  • Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

Relative Effect of Glyphosate on Glyphosate-Tolerant Maize Rhizobacterial Communities is Not Altered by Soil Properties

  • Barriuso, Jorge;Mellado, Rafael P.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.159-165
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    • 2012
  • The rhizobacterial composition varies according to the soil properties. To test if the effect of herbicides on the rhizobacterial communities of genetically modified NK603 glyphosate-tolerant maize varies according to different soil locations, a comparison was made between the effects of glyphosate (Roundup Plus), a post-emergence applied herbicide, and a pre-emergence applied herbicide (GTZ) versus untreated soil. The potential effect was monitored by direct amplification, cloning, and sequencing of the soil DNA encoding 16S rRNA, and high-throughput DNA pyrosequencing of the bacterial DNA coding for the 16S rRNA hypervariable V6 region. The results obtained using three different methods to analyze the herbicide effect on the rhizobacterial communities of genetically modified NK603 maize were comparable to those previously obtained when glyphosate-tolerant maize was grown in soil with different characteristics. Both herbicides decreased the bacterial diversity in the rhizosphere, with Actinobacteria being the taxonomic group most affected. The results suggest that both herbicides affected the structure of the maize rhizobacterial community, but glyphosate was environmentally less aggressive.

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.174-179
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    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Programmed Cell Death in Bacterial Community: Mechanisms of Action, Causes and Consequences

  • Lee, Heejeong;Lee, Dong Gun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1014-1021
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    • 2019
  • In the bacterial community, unicellular organisms act together as a multicellular being. Bacteria interact within the community and programmed cell death (PCD) in prokaryotes is a sort of altruistic action that enables the whole population to thrive. Genetically, encoded cell death pathways are triggered by DNA damage or nutrient starvation. Given the environmental and bacterial diversity, different PCD mechanisms are operated. Still, their biochemical and physiological aspects remain unrevealed. There are three main pathways; thymineless death, apoptosis-like death, and toxin-antitoxin systems. The discovery of PCD in bacteria has revealed the possibility of developing new antibiotics. In this review, the molecular and physiological characteristics of the three types of PCD and their development potential as antibacterial agents are addressed.

CpG DNA for Treatment of Allergic Diseases (알레르기 질환의 치료로서의 CpG DNA)

  • Choi, Sung Min
    • Clinical and Experimental Pediatrics
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    • v.48 no.3
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    • pp.251-259
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    • 2005
  • Atopy is a highly prevalent and serious health problem. The prevalence and severity of asthma and allergic diseases have increased over recent decades, particularly in industrialized nations. Early life infections may protect against the development of atopy and allergic diseases like asthma. The inverse relationship between the incidence of atopy and childhood infections has led to the 'hygiene hypothesis', which suggests that diminished exposure to childhood infections in modern society has led to decreased Th1-type responses. Th1 and Th2 responses are counter-regulatory. Reduced Th1 may lead to enhanced Th2-type inflammation, which is important in promoting asthma and allergic disease via up-regulation of IL-4, IL-5, and IL-13. It is now widely accepted that altered regulation of Th2 responses(and possibly the balance between Th1 and Th2 responses) is an important factor in the development of atopy. CpG DNA represent a novel class of drugs with substantial immunomodulatory properties. CpG DNA contain unmethylated motifs centered on the CpG dinucleotides, like bacterial DNA. These CpG DNA promote Th1 and regulatory type immune responses and suppress Th2 responses. In murine studies, CpG DNA are effective in prevention and treatment of asthma and allergic diseases. CpG DNA are just beginning to be tested in human asthma. While its precise mechanisms continue to be fully studied, CpG DNA offers considerable promise as a novel treatment for atopic inflammation. It may prove to be an important disease modifying therapy, or even curative therapeutic agent for asthma and allergic diseases.