• Title/Summary/Keyword: M-DNA

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Branched DNA-based Synthesis of Fluorescent Silver Nanocluster

  • Park, Juwon;Song, Jaejung;Park, Joonhyuck;Park, Nokyoung;Kim, Sungjee
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1105-1109
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    • 2014
  • While single strand DNAs have been widely used for the scaffold of brightly fluorescent silver nanoclusters (Ag NCs), double strand DNAs have not been as successful. Herein, we report a novel synthetic approach for bright Ag NCs using branched double strand DNAs as the scaffolds for synthesis. X-shaped DNA (X-DNA) and Y-shaped DNA (Y-DNA) effectively stabilized Ag NCs, and both X-DNA and Y-DNA resulted in brightly fluorescent Ag NCs. The concentration and molar ratio of silver and DNA were found important for the fluorescence efficiency. The brightest Ag NC with the photoluminescence quantum efficiency of 19.8% was obtained for the reaction condition of 10 ${\mu}M$ X-DNA, 70 ${\mu}M$ silver, and the reaction time of 48 h. The fluorescence lifetime was about 2 ns for the Ag NCs and was also slightly dependent on the synthetic condition. Addition of Cu ions at the Ag NC preparations resulted in the quenching of Ag NC fluorescence, which was different to the brightening cases of single strand DNA stabilized Ag NCs.

Transformation of Bacillus brevis P176-2 with Plasmid DNA by Electroporation (전기천공법에 의한 Bacillus brevis P176-2의 형질전환)

  • 채기수;엄경일
    • The Korean Journal of Food And Nutrition
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    • v.5 no.2
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    • pp.77-83
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    • 1992
  • The optimum conditions and mechanisms for the plasmid-mediated genetic transformation of intact cells of Bacillus brevis Pl76-2, an extracellular protein producing bacterium by electroporation were investigated. It was found that pUB110 Plasmid DNA can be introduced into intact bacterial cells by electroporation. The frequency of transformation by this electroporation system depended upon the initial electric field strength, the capacity of the electric discharge capacitor, growth stage, number of successive pulses and composition of electroporation buffer. It was effective for transformation that cells were harvested, washed and resuspended with HSM [7M HEPES(PH 7.4), 272mM sucrose, 1 mM MgCl2] electroporation buffer when cell growth was attained to 1.2 at OD660. A maximum frequency of transformation of 2.40$\times$104 transformants per$\mu$g plasmid DNA was obtained by two succesive Pulses with an initial electric field strength of 12.5kV/cm and with a capacitance of 7.3uF.

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Protective Effects of Ginkgo Biloba Leaf Extract(GBE) against 1,2,4-benzenetriol Induced Toxicity in Vitro (Ginkgo biloga 잎 추출물의 1,2,4-benzenetriol에 대한 항산화 효과에 대한 연구)

  • 이영준;김태연;정해원
    • Journal of Environmental Health Sciences
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    • v.27 no.1
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    • pp.124-130
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    • 2001
  • Ginkgo biliba has been used for bronchitis and asthma in oriental countries and its leaf extract(GBE) contains 24% ginkgoflavone glycoside and 6% terpenoid. Flavonoids and terpenoids are known to have various antioxidant effects such as scavenging of free radicals and chelation of transtional metals. Antioxidant effect of GBE against 1,2,4-benzenetriol(BT), one of toxic metabolites of benzene, was demonstrated throughbsister chromatid exchange(SCE) analysis, single cell gel electrophoresis(SCGE) analysis, DNA cleavage assay and lipid peroxidation production analysis. The means of SCE frequencies at 10, 25 and 50$\mu$M concentration of BT were 7.72, 8.02, 9.22 respectively. In addition of GBE with concentration of 50, 200 and 500$\mu\textrm{g}$/$m\ell$, SCE frequencies were decreased significantly.(p<0.05) According to SCGE analysis, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 $\mu$m and the DNA damage induced by BT was significantly protected by GBE(p<0.001). No genotoxicity was observed by GBE treatment alone on DNA cleavage. The effect of BT on lipid peroxidation product, Malondiadehyde(MDA), was increased with concentration of BT(10 and 50 $\mu$M) and reduction in MDA was noted when GBE was added. From above results it is suggested that GBE could protect the cell and DNA from pro-oxidant effect by reactive oxigen species induced by BT.

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Expression of Poly (ADP-ribose) Polymerase During Apoptosis Induced by Ultraviolet Radiation in HeLa $S_3$ Cells

  • Chang, Jeong-Hyun;Kwon, Heun-Young
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.349-354
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    • 2006
  • Induction of apoptosis allows the organism to get rid of abnormal cells and also of tumor cells. Understanding the mechanism involved in Ultraviolet radiation (UV) induced apoptosis may improve its therapeutic efficacy. In this study, we present expression of poly (ADP-ribose) polymerase (PARP) during apoptosis induced by UV in HeLa $S_3$ cells. Four different assays were performed in this study: morphological assessment of apoptotic cells and cell viability, DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, and expression of PARP by the western blot analysis. The percentages of apoptotic HeLa $S_3$ cells irradiated with $75J/m^2$ UV was increased continuously from 3 hrs incubation. DNA ladder pattern was appeared at 6 hrs. The amount of nucleosomal DNA fragments in cells treated UV increased from 3 to 12 hrs incubation and gradually decreased. The cleavage of PARP in HeLa $S_3$ cells irradiated with UV was induced, and the cleavage of PARP was more delayed in the cells pretreated with $5J/m^2$ UV and subsequently irradiated with $75J/m^2$ UV. than that in the cells only irradiated with $75J/m^2$ UV. Thus these data suggest that the cleavage of PARP relates with DNA fragmentation associated with apoptosis.

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Analysis of partial cDNA sequence from Theileria sergenti

  • Park, Jin-ho;Chae, Joon-seok;Kim, Dae-hyuk;Jang, Yong-suk;Kwon, Oh-deog;Lee, Joo-mook
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.797-805
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    • 1999
  • T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

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Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle

  • Baik, M.;Vu, T.T.T.;Piao, M.Y.;Kang, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1493-1498
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    • 2014
  • Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

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.

Cytogenetic Study of Maackia amurensis Rupr. & Maxim. and M. fauriei (Levl.) Takeda Using Karyotyping Analysis and the FISH Technique (핵형분석과 FISH 기술을 이용한 솔비나무와 다릅나무의 세포유전학적 연구)

  • Kim, Soo-Young;Kim, Chan-Soo
    • Korean Journal of Plant Taxonomy
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    • v.39 no.3
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    • pp.193-198
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    • 2009
  • Chromosome analysis using karyotyping and bicolor FISH were carried out for two Maackia species (M. fauriei and M. amurensis) found in Korea. The somatic metaphase chromosome number was 2n = 2x = 18 in both, and the size of these chromosomes ranged from 3.58 to $5.82{\mu}m$. The chromosome complements consisted of two pairs of metacentric (chromosomes 1 and 7), four pairs of submetacentrics (chromosomes 4, 6, 8 and 9) and three pairs of subtelocentrics (chromosomes 2, 3 and 5) in M. fauriei but, chromosomes 4 (subtelocentric) and 7 (submetacentric) of M. amurensis have different morphology. Using bicolor FISH, a pair of 45S rDNA loci were observed for both M. fauriei and M. amurensis, but the number and site of the 5S rDNA signal were different in the two species. M. fauriei has two pairs of 5S signals on chromosomes 7 and 8 but, M. amurensis has four paris on chromosomes 3, 4, 7 and 7. Hence, the 5S rDNA is a useful FISH for Maackia species.

Comparison and Evaluation on the Chemical Constituents of Progeny in T-DNA Inserted Rice

  • Qin, Yang;Kim, Suk-Man;An, Gyn-Heung;Sohn, Jae-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.131-136
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    • 2008
  • With the development of diverse agricultures worldwide, biofortified rice noted for its preferable marketability and palatability plays an important role in the world's agricultural economics and rice breeding programs. In this report, several $M_5$ of T-DNA inserted lines derived from the donor cultivars, 'Hwayong' and 'Dongjin', were selected for high or low protein, high lipid and low amylose content, respectively. The coefficients and ranges of variation for the chemical constituents between $M_4$ and $M_5$ T-DNA inserted lines were evaluated in comparison with those of the donor varieties. Results indicated that T-DNA insertion might be an effective way to generate useful variations for chemical composition of rice grains which could be used for the development of biofortified rice cultivars.

A Simple and Reliable Method for Preparation of Cross-Contamination-Free Plant Genomic DNA for PCR-Based Detection of Transgenes

  • Hwang, Seon-Kap;Kim, Young-Mi
    • BMB Reports
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    • v.33 no.6
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    • pp.537-540
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    • 2000
  • A simplified but reliable method was developed for the polymerase chain reaction (PCR)-based detection of genetically modified (GM) plants. The modified CTAB (mCTAB) method enabled us to prepare a high quality of genomic DNA from several hundred plant leaf samples in one day. Using DNA samples prepared from seven dicots and two monocots, approximately 1.75-kb regions spanning 17 S to 25 S ribosomal RNA genes were successfully amplified in a 2X PCR pre-mix containing BLOTTO. Further fidelity assessment of the mCTAB method by PCR analysis with Roundup Ready soybean (RRS) and non-RRS plants showed that the DNA samples prepared alternately from each of two lines were evidently free of cross-contamination. These results demonstrate that the mCTAB method is highly recommended for the rapid detection of transgenes in large numbers of leaf samples from diverse transgenic plants.

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