• Title/Summary/Keyword: environmental DNA (eDNA)

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REP-PCR Genotyping of Four Major Gram-negative Foodborne Bacterial Pathogens (주요 식중독 그람 음성 세균 4속의 REP-PCR genotyping)

  • Jung, Hye-Jin;Seo, Hyeon-A;Kim, Young-Joon;Cho, Joon-Il;Kim, Keun-Sung
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.611-617
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    • 2005
  • Dispersed repetitive DNA elements in genomes of microorganisms differ among and within species. Because distances between repetitive sequences vary depending on bacterial strains, genomic fingerprinting with interspersed repetitive sequence-based probes can be used to distinguish unrelated organisms. Among well-known bacterial repetitive sequences, Repetitive Extragenic Palindromic (REP) sequence has been used to identify environmental bacterial species and strains. We applied REP-PCR to detect and differentiate four major Gram-negative food-borne bacterial pathogens, E. coli, Salmonella, Shigella, and Vibrio. Target DNA fragments of these pathogens were amplified by REP-PCR method. PCR-generated DNA fragments were separated on 1.5% agarose gel. Dendrograms for PCR products of each strain were constructed using photo-documentation system. REP-PCR reactions with primer pairs REP1R-I and REP2-I revealed distinct REP-PCR-derived genomic fingerprinting patterns from E. coli, Salmonella, Shigella, and Vibrio. REP-PCR method provided clear distinctions among different bacterial species containing REP-repetitive elements and can be widely used for typing food-borne Gram-negative strains. Results showed established REP-PCR reaction conditions and generated dendrograms could be used with other supplementary genotyping or phenotyping methods to identify isolates from outbreak and to estimate relative degrees of genetic similarities among isolates from different outbreaks to determine whether they are clonally related.

A Molecular Biotechnology For Removal of Toxic Heavy Metals

  • Bang Sang-Weon;Clark Douglas S.;Keasling Jay D.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.128-135
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    • 2000
  • The thiosulfate reductase gene (PhsABC) from Salmonella typhimurium was expressed in Escherichia coli in order to produce sulfide from inorganic thiosulfate and precipitate metals as metal sulfide complexes. A 5.1-kb DNA fragment containing the native phsABC and a 3.7-kb DNA fragment, excluding putative promoter and regulatory regions were inserted into expression vectors pTrc99A and pJB866, respectively. Upon expression of phsABC, E. coli DH5$\alpha$ harboring the phsABC constructs showed higher thiosulfate reductase activity and produced significantly more sulfide than the control strain (E. coli DH5$\alpha$) under both aerobic and anaerobic conditions. Among the four constructs, E. coli DH5$\alpha$ harboring pSB74 produced the highest level of thiosulfate reductase and removed most of heavy metals from solution under anaerobic conditions. In a mixture of 100 $\mu$M each of cadmium, lead, and zinc, the strain could remove $99\%$ of the total metals from solution within 10 hours. Cadmium was removed first, lead second, and zinc last. In contrast, a negative control did not produce any measurable sulfide and removed very little metals from solution. These results have important implications for removal of metals from wastewater contaminated with several metals.

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Earthworm Enhanced Bioaugmentation of PCB Contaminated Soil

  • Crowley, David E.;Luepromchai, Ekawan;Singer, Andrew S.;Yang, Chang Sool
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.100-107
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    • 2000
  • In a recently developed strategy for in-situ treatment of polychlorinated biphenyls (PCB), bioaugmentation was used in conjunction with a surfactant, sorbitan trioleate, as a carbon source for the degrader bacteria, along with the monoterpene, carvone, and salicylic acid as inducing substrates. Two bacteria were used for soil inoculants, including Arthrobacter sp. st. B1B and Ralstonia eutrophus H850. This methodology achieved 60% degradation of PCBs in Aroclor 1242 after 18 weeks in soils receiving 34 repeated applications of the degrader bacteria. However, an obvious limitation was the requirement for soil mixing after every soil inoculation. In the research reported here, bioaugmentation and biostimulation treatment strategies were modified by using the earthworm, Pheretima hawayana, as a vector for dispersal and mixing of surface-applied PCB-degrading bacteria and soil chemical amendments. Changes in microbial biomass and microbial community structure due to earthworm effects were examined using DNA extraction and PCR-DGGE of 16S rDNA. Results showed that earthworms effectively promoted biodegradation of PCBs in bioaugmented soils to the same extent previously achieved using physical soil mixing, and had a lesser, but significant effect in promoting PCB biodegradation in biostimulated soils treated with carvone and salicylic acid. The effects of earthworms were speculated to involve many interacting factors including increased bacterial transport to lower soil depths, improved soil aeration, and enhanced microbial activity and diversity.

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Comparative Viral Metagenomics of Environmental Samples from Korea

  • Kim, Min-Soo;Whon, Tae Woong;Bae, Jin-Woo
    • Genomics & Informatics
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    • v.11 no.3
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    • pp.121-128
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    • 2013
  • The introduction of metagenomics into the field of virology has facilitated the exploration of viral communities in various natural habitats. Understanding the viral ecology of a variety of sample types throughout the biosphere is important per se, but it also has potential applications in clinical and diagnostic virology. However, the procedures used by viral metagenomics may produce technical errors, such as amplification bias, while public viral databases are very limited, which may hamper the determination of the viral diversity in samples. This review considers the current state of viral metagenomics, based on examples from Korean viral metagenomic studies-i.e., rice paddy soil, fermented foods, human gut, seawater, and the near-surface atmosphere. Viral metagenomics has become widespread due to various methodological developments, and much attention has been focused on studies that consider the intrinsic role of viruses that interact with their hosts.

Response of Bioluminescent Bacteria to Sixteen Azo Dyes

  • Lee, Hwa-Young;Park, Sue-Hyung;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.101-105
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    • 2003
  • Recombinant bioluminescent bacteria were used to monitor and classify the to xicity of azo dyes. Two constitutive bioluminescent bacteria, Photobacterium phosphoreum and Es-Cherichia coli, E, coli GC2 (lac::luxCOABE), were used to detect the cellular toxicity of the azo dyes. In addition, four stress-inducible bioluminestent E. coli, DPD2794 (recA::luxCDABE), a DNA damage Sensitive strain; DPD2540 (fabA::luxCDABE), a membrane damage sensitive strain; DPD2511 (katG::luxCDABE), an oxidative damage sensitive strain; and TV1061 (grpE::luxCDABE), a protein damage sensitive strain, were used to provide information about the type of toxicity caused by crystal violet, the most toxic dye of the 16 azo dyes tested. These results suggest that azo dyes result in serious cellular toxicity in bacteria, and that toxicity monitoring and classific ation of some azo dyes, In the field, may be possible using these recombinant bioluminescent bacteria.

Occurrence of Powdery Mildew Caused by Erysiphe abeliicola on Glossy Abelia in Korea (Erysiphe abeliicola에 의한 꽃댕강나무 흰가루병 발생)

  • Cho, Sung-Eun;Park, Ji-Hyun;Lee, Seung-Kyu;Lee, Sang-Hyun;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.18 no.2
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    • pp.133-138
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    • 2012
  • In November 2009, a powdery mildew on glossy abelia (Abelia ${\times}$ grandiflora) was found in Seogwipo, Jeju Island, Korea. Further survey in the southern part of Korea, e.g., Jeju, Busan, and Tongyeong confirmed occurrence of the disease. White colonies were present on leaves, young stems, and flowers, detracting from their beauty in landscape plantings. Severely infected lesions were discolored to red-purplish. Based on the morphological characteristics and analysis of rDNA, the fungus associated with the symptoms was identified as Erysiphe abeliicola U. Braun & S. Takam. This work provides the morphological feature of its anamorph for the first time, which is characterized by having multi-lobed hyphal appressoria and short foot-cells of conidiophores. Morphological characteristics of mature chasmothecia were consistent with the previous Japanese record of this species. The sequence of internal transcribed spacer region of ribosomal DNA obtained from a Korean sample showed that this species places in the section Microsphaera of the genus Erysiphe in phylogenetic position, corresponding with the classical taxonomy. This is the first report of E. abeliicola and its host plant in Korea. The host plant A. ${\times}$ grandiflora is newly listed in the host range of E. abeliicola.

The first review study on association of DNA methylation with gastric cancer in Iranian population

  • Shahbazi, Mahsa;Yari, Kheirollah;Rezania, Niloufar
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.5
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    • pp.2499-2506
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    • 2016
  • Background: Gastric cancer (GC) is the second leading cause of cancer-related death worldwide. Several environmental, genetic and epigenetic factors have been suggested to have a role in GC development. Epigenetic mechanisms like histone changes and promoter hyper-methylation are now being increasingly studied. Associations between methylation of many gene promoters with the risk of gastric cancer have been investigated worldwide. Such aberrant methylation may result in silencing of specific genes related to cell cycling, cell adhesion, apoptosis and DNA repair. Thus this molecular mechanism might have a key role in proliferation and migration of cancerous cells. Materials and Methods: In this review article we included studies conducted on DNA methylation and gastric cancer in Iranian populations. Using Science direct, Pubmed/PMC, Springer, Wiley online library and SciELO databases, all published data until 31 January 2016 were gathered. We also searched Science direct data base for similar investigations around the world to make a comparison between Iran and other countries. Results: By searching these databases, we found that the association between methylation of seven gene promoters and gastric cancer had been studied in Iran until 31 January 2016. These genes were p16, hLMH1, E-cadherin, CTLA4, $THR{\beta}$, mir9 and APC. Searching in science direct database also showed that 92 articles had been published around the world till January 2016. Our investigation revealed that despite the importance of GC and its high prevalence in Iran, the methylation status of only a few gene promoters has been studied so far. More studies with higher sample numbers are needed to reveal the relation of methylation status of gene promoters to gastric cancer in Iran. Conclusions: Further studies will be helpful in identifying associations of DNA methylation in candidate genes with gastric cancer risk in Iranian populations.

Monitoring of Geothermal Systems Wells and Surrounding Area using Molecular Biological Methods for Microbial Species (분자생물학적 방법을 이용한 지열시스템 관정 및 주변지역 미생물종 모니터링)

  • Ahn, Chang-Min;Han, Ji-Sun;Kim, Chang-Gyun;Park, Yu-Chul;Mok, Jong-Koo;Jang, Bum-Ju
    • Journal of Soil and Groundwater Environment
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    • v.17 no.6
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    • pp.23-32
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    • 2012
  • This study was conducted to monitor microbial species dynamics within the aquifer due to long term operation of geothermal heat pump system. The species were identified by molecular biological methods of 16S rDNA. Groundwater sample was collected from both open (S region) and closed geothermal recovery system (J region) along with the control. J measured and control as well as S measured found Ralstonia pickettii as dominant species at year 2010. In contrast, Rhodoferax ferrireducens was dominantly observed for the control of S. In 2011, Sediminibacterium sp. was universely identified as the dominant species regardless of the monitoring places and type of sample, i.e., measured or control. The difference in the dynamics between the measured and the control was not critically observed, but annual variation was more strikingly found. It reveals that possible environmental changes (e.g. ORP and DO) due to the operation of geothermal heat recovery system in aquifer could be more exceedingly preceded to differentiate annual variation of microbial species rather than positional differences.

Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

  • Maeng, Chang-Jae;Kim, Hyung-Kwoun;Park, Sun-Yang;Koo, Bon-Tag;Oh, Tae-Kwang;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.85-91
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    • 2001
  • To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

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