• Title/Summary/Keyword: DNA persistence

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In Vivo Kinetics and Biodistribution of a HIV-1 DNA Vaccine after Administration in Mice

  • Kim, Byong-Moon;Lee, Dong-Sop;Choi, Jae-Hoon;Kim, Chae-Young;Son, Mi-Won;Suh, You-Suk;Baek, Kwan-Hyuck;Park, Ki-Seok;Sung, Young-Chul;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • v.26 no.6
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    • pp.493-498
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    • 2003
  • In this study we have investigated the pharmacokinetics and tissue distribution of GX-12, a multiple plasmid DNA vaccine for the treatment of HIV-1 infection. Plasmid DNA was rapidly degraded in blood with a half-life of 1.34 min and was no longer detectable at 90 min after intravenous injection in mice. After intramuscular injection, plasmid DNA concentration in the injection site rapidly declined to less than 1 % of the initial concentration by 90 min post-injection. However, sub-picogram levels (per mg tissue) were occasionally detected for several days after injection. The relative proportions of the individual plasm ids of GX-12 remained relatively constant at the injection site until 90 min post-injection. The concentration of plasmid DNA in tissues other than the injection site peaked at 90 min post-injection and decreased to undetectable levels at 8 h post-injection. The rapid in vivo degradation of GX-12 and absence of persistence in non-target tissues suggest that the risk of potential gene-related toxicities by GX-12 administration, such as expression in non-target tissues, insertional mutagenesis and germline transmission, is minimal.

Effect of novel luminol-based blood detection reagents on DNA stability (새로운 루미놀 기반 혈흔 탐지 시약이 디엔에이에 미치는 영향에 대한 연구)

  • Jung, Ju Yeon;Oh, Yu-Li;Lee, Jee Won;Lim, Seung;Kim, Jung-mok;Lee, Yang Han;Lim, Si-Keun
    • Analytical Science and Technology
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    • v.31 no.2
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    • pp.71-77
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    • 2018
  • Detection of bloodstains is a very important process in scientific investigations, and luminol is often used for the detection of bloodstains that are not visible. Recently, new preparation methods of blood detection reagents based on luminol (BloodFlareA, B) were developed and reported to have higher active persistence and to be more economical than conventional blood detection reagent, BlueStar forensic. In this paper, we tested the specificity and effect of the BloodFlares (A and B) on DNA and compared them with those of BlueStar forensic. False positive results for the BloodFlares were not observed in semen, saliva, vaginal fluids, urine, sweat, and nasal discharge, but were observed in $CuSO_4$, $FeSO_4$, and bleach solutions, and the observed patterns were similar to those of BlueStar forensic. The effect on DNA was determined by analyzing the DNA yield, degradation index, and DNA profiling. Based on these results, we concluded that the BloodFlares based on luminol do not affect DNA stability and are applicable in forensics.

Contamination patterns of Listeria spp. in pork processing plants using random amplified polymorphic DNA (RAPD를 이용한 돈육 가공장의 Listeria 오염양상 분석)

  • Ha, Sung-Yeol;Choi, Weon-Sang;Bahk, Gyung-Jin;Hong, Chong-Hae
    • Korean Journal of Veterinary Research
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    • v.45 no.3
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    • pp.359-367
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    • 2005
  • This study was carried out to understand the contamination patterns of Listeria in pork processing plants. A total of 402 samples were collected from carcass, pork during processing, surfaces of equipment and environment, and 238 isolates of Listeria species were identified. L. innocua was found in 64.7% of the isolates, L. monocytogenes in 33.2%, and L. welshimeri in 2.1%. Random amplified polymorphic DNA (RAPD) analysis performed to investigate the origin and routes of Listeria contamination, showed 21 composite types of L. monocytogenes and 26 composite types of L. innocua. It was confirmed that Listeria contamination begins with contaminated incoming carcass and ever-present contaminants in the processing environments. The persistence and dissemination of the same strain of L. monocytogenes and L. innocua throughout the processing line revealed that the sanitation standard operating procedure should be implemented to minimize the risk of colonization in the workplace. Molecular subtyping of L. innocua allowed us to tracing the possibility of cross-contamination during processing.

Revealing Joseon period People's single nucleotide polymorphism associated with lactase gene by ancient DNA analysis of human remains from archaeological sites in Korea

  • Chang Seok Oh;Myeung Ju Kim;Yi-Suk Kim;Sori Min;Kyong Taek Oh;Soong Deok Lee;Dong Hoon Shin
    • Anatomy and Cell Biology
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    • v.56 no.1
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    • pp.54-60
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    • 2023
  • Lactase non-persistence (LNP), one of the causes of lactose intolerance, is related to lactase gene associated single nucleotide polymorphisms (SNPs). Since the frequency of LNP varies by ethnic group and country, the research to reveal the presence or absence of LNP for specific people has been conducted worldwide. However, in East Asia, the study of lactase gene associated SNPs have not been sufficiently examined so far using ancient human specimens from archaeological sites. In our study of Joseon period human remains (n=14), we successfully revealed genetic information of lactase gene associated SNPs (rs1679771596, rs41525747, rs4988236, rs4988235, rs41380347, rs869051967, rs145946881 and rs182549), further confirming that as for eight SNPs, the pre-modern Korean people had a lactase non-persistent genotype. Our report contributes to the establishment of LNP associated SNP analysis technique that can be useful in forthcoming studies on human bones and mummy samples from East Asian archaeological sites.

Persistence of Multidrug-Resistant Acinetobacter baumannii Isolates Harboring blaOXA-23 and bap for 5 Years

  • Sung, Ji Youn;Koo, Sun Hoe;Kim, Semi;Kwon, Gye Cheol
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1481-1489
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    • 2016
  • The emergence and dissemination of carbapenemase-producing Acinetobacter baumannii isolates have been reported worldwide, and A. baumannii isolates harboring blaOXA-23 are often resistant to various antimicrobial agents. Antimicrobial resistance can be particularly strong for biofilm-forming A. baumannii isolates. We investigated the genetic basis for carbapenem resistance and biofilm-forming ability of multidrug-resistant (MDR) clinical isolates. Ninety-two MDR A. baumannii isolates were collected from one university hospital located in the Chungcheong area of Korea over a 5-year period. Multiplex PCR and DNA sequencing were performed to characterize carbapenemase and bap genes. Clonal characteristics were analyzed using REP-PCR. In addition, imaging and quantification of biofilms were performed using a crystal violet assay. All 92 MDR A. baumannii isolates involved in our study contained the blaOXA-23 and bap genes. The average absorbance of biomass in Bap-producing strains was much greater than that in non-Bap-producing strains. In our study, only three REP-PCR types were found, and the isolates showing type A or type B were found more than 60 times among unique patients during the 5 years of surveillance. These results suggest that the isolates have persisted and colonized for 5 years, and biofilm formation ability has been responsible for their persistence and colonization.

Effect of Producing Different Phenazines on Bacterial Fitness and Biological Control in Pseudomonas chlororaphis 30-84

  • Yu, Jun Myoung;Wang, Dongping;Pierson, Leland S. III;Pierson, Elizabeth A.
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.44-58
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    • 2018
  • Pseudomonas chlororaphis 30-84 is a biological control agent selected for its ability to suppress diseases caused by fungal pathogens. P. chlororaphis 30-84 produces three phenazines: phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine-1-carboxylic acid (2OHPCA) and a small amount of 2-hydroxy-phenazine (2OHPHZ), and these are required for fungal pathogen inhibition and wheat rhizosphere competence. The two, 2-hydroxy derivatives are produced from PCA via the activity of a phenazine-modifying enzyme encoded by phzO. In addition to the seven biosynthetic genes responsible for the production of PCA, many other Pseudomonas strains possess one or more modifying genes, which encode enzymes that act independently or together to convert PCA into other phenazine derivatives. In order to understand the fitness effects of producing different phenazines, we constructed isogenic derivatives of P. chlororaphis 30-84 that differed only in the type of phenazines produced. Altering the type of phenazines produced by P. chlororaphis 30-84 enhanced the spectrum of fungal pathogens inhibited and altered the degree of take-all disease suppression. These strains also differed in their ability to promote extracellular DNA release, which may contribute to the observed differences in the amount of biofilm produced. All derivatives were equally important for survival over repeated plant/harvest cycles, indicating that the type of phenazines produced is less important for persistence in the wheat rhizosphere than whether or not cells produce phenazines. These findings provide a better understanding of the effects of different phenazines on functions important for biological control activity with implications for applications that rely on introduced or native phenazine producing populations.

Partial Reduction of Dinitroaniline Herbicide Pendimethalin by Bacillus sp. MS202 (Bacillus sp. MS202에 의한 Dinitroaniline계 제초제인 Pendimethalin의 부분환원)

  • Lee, Young-Keun;Chang, Hwa-Hyoung;Jang, Yu-Sin;Hyung, Seok-Won;Chung, Hye-Young
    • Korean Journal of Environmental Agriculture
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    • v.23 no.4
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    • pp.197-202
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    • 2004
  • The persistence of pendimethalin in soil and ground water has an injurious effect on ecosystem. Pendimethalin-degrading bacterium was isolated from Masan, Gyeongnam province and temporarily identified as Bacillus sp. MS202 by the analysis of API CHB50, kit, FAME, and 16S rDNA sequence. from the analysis of pnedimethalin metabolite using TLC, GC, and GC-MS, we found that the degradation of pendimethalin by Bacillus sp. MS202 did not result in the dealkylated form, but the formation of the reduced compound, 6-amino-2-nitro-N(1-ethylpropyl)-3,4-xylidine or 2- amino-6-nitro-N(1-ethylpropyl)-3,4-xylidine.

Recurrent Pseudomonas aeruginosa Infection in Chronic Lung Diseases: Relapse or Reinfection?

  • Yum, Ho-Kee;Park, I-Nae;Shin, Bo-Mun;Choi, Soo-Jeon
    • Tuberculosis and Respiratory Diseases
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    • v.77 no.4
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    • pp.172-177
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    • 2014
  • Background: Pseudomonas aeruginosa infection is particularly associated with progressive and ultimately chronic recurrent respiratory infections in chronic obstructive pulmonary disease, bronchiectasis, chronic destroyed lung disease, and cystic fibrosis. Its treatment is also very complex because of drug resistance and recurrence. Methods: Forty eight cultures from 18 patients with recurrent P. aeruginosa pneumonia from 1998 to 2002 were included in this study. Two or more pairs of sputum cultures were performed during 2 or more different periods of recurrences. The comparison of strains was made according to the phenotypic patterns of antibiotic resistance and chromosomal fingerprinting by pulsed field gel electrophoresis (PFGE) using the genomic DNA of P. aeruginosa from the sputum culture. Results: Phenotypic patterns of antibiotic resistance of P. aeruginosa were not correlated with their prior antibiotic exposition. Fifteen of 18 patients (83.3%) had recurrent P. aeruginosa pneumonia caused by the strains with same PFGE pattern. Conclusion: These data suggest that the most of the recurrent P. aeruginosa infections in chronic lung disease occurred due to the relapse of prior infections. Further investigations should be performed for assessing the molecular mechanisms of the persistent colonization and for determining how to eradicate clonal persistence of P. aeruginosa.

Microbial linguistics: perspectives and applications of microbial cell-to-cell communication

  • Mitchell, Robert J.;Lee, Sung-Kuk;Kim, Tae-Sung;Ghim, Cheol-Min
    • BMB Reports
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    • v.44 no.1
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    • pp.1-10
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    • 2011
  • Inter-cellular communication via diffusible small molecules is a defining character not only of multicellular forms of life but also of single-celled organisms. A large number of bacterial genes are regulated by the change of chemical milieu mediated by the local population density of its own species or others. The cell density-dependent "autoinducer" molecules regulate the expression of those genes involved in genetic competence, biofilm formation and persistence, virulence, sporulation, bioluminescence, antibiotic production, and many others. Recent innovations in recombinant DNA technology and micro-/nano-fluidics systems render the genetic circuitry responsible for cell-to-cell communication feasible to and malleable via synthetic biological approaches. Here we review the current understanding of the molecular biology of bacterial intercellular communication and the novel experimental protocols and platforms used to investigate this phenomenon. A particular emphasis is given to the genetic regulatory circuits that provide the standard building blocks which constitute the syntax of the biochemical communication network. Thus, this review gives focus to the engineering principles necessary for rewiring bacterial chemo-communication for various applications, ranging from population-level gene expression control to the study of host-pathogen interactions.

Impact of Virus-resistant Trigonal Cactus Cultivation on Soil Microbial Community (바이러스저항성 삼각주 재배가 토양 미생물상에 미치는 영향)

  • Oh, Sung-Dug;Kim, Jong-Bum;Lee, Jung-Jin;Kim, Min-Kyeong;Ahn, Byung-Ohg;Sohn, Soo-In;Park, Jong-Sug;Ryu, Tae-Hun;Cho, Hyun-Suk;Lee, Kijong
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.148-154
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    • 2013
  • BACKGROUND: Genetically modified(GM) trigonal cactus(Hylocereus trigonus Saff.) contained a coat protein gene of cactus virus X (CVX), which conferred resistance to the virus, phosphinothricin acetyltransferase (bar) gene, which conferred herbicide resistance, and a cauliflower mosaic virus 35S promoter (CaMV 35S). This study was conducted to evaluate the possible impact of GM trigonal cactus cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM trigonal cactus cultivation soils. The total numbers of bacteria, and actinomycete in the rhizosphere soils cultivated GM and non-GM trigonal cactus were similar to each other, and there was no significant difference. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM trigonal cactus were Proteobacteria, Uncultured archaeon, and Uncultured bacterium. The denaturing gradient gel electrophoresis (DGGE) profiles show a similar patterns, significant difference was not observed in each other. DNA was isolated from soil cultivated GM and non-GM trigonal cactus, we analyzed the persistence of the inserted gene by PCR. Amplification of the inserted genes was not observed in the soil DNA, which was collected after harvest. CONCLUSION(S): This result suggests that the GM trigonal cactus cultivation does not change significantly the microbial community.