• Title/Summary/Keyword: M-DNA

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Differentiation of four Mycobacterium Species using DNA-DNA Hybridization Method using Specific Probes

  • Kweon, Tae-Dong;Bai, Sun-Joon;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2013.05a
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    • pp.1012-1014
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    • 2013
  • DNA-DNA hybridization method with four oligonucleotide-specific probes was used simultaneously for differentiation and identification of four Mycobacterium species (Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii). This DNA-DNA hybridization method with 4 oligonucleotide-specific probes, which targets in the rpoB region of 4 Mycobacteria species, respectively, was tested on 322 clinical isolates. Using DNA-DNA hybridization method, we detected M. tuberculosis (282 strains), M. avim (7 strains), M. intracellulare (9 strains), and M. kansasii (3 strain) from 322 clinical isolates. This result was compared with conventional biochemical test and rpoB DNA sequence analysis of this clinical isolates. We confirmed identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii with high sensitivity (100 %) and specificity (100 %). This DNA-DNA hybridization method could be performed within 4 hours at least. Therefore, we suggest that DNA- DNA hybridization method using 4 rpoB DNA probes of Mycobacteria could be used for accurate, rapid, convenient detection and identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii in clinical samples.

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Sensitivity of a charge-detecting label-free DNA sensor using field-effect transistors (FETs) depending on the Debye length (전계효과 트랜지스터(FETs)를 이용한 전하 검출형 DNA 센서에서 Debye length에 따른 검출 감도)

  • Song, Kwang-Soup
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.48 no.2
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    • pp.86-90
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    • 2011
  • The effects of cations are very important in field-effect transistors (FETs) type DNA sensors detecting the intrinsic negative charge between single-stranded DNA and double-stranded DNA without labeling, because the intrinsic negative charge of DNA is neutralized by cations in electrolyte solution. We consider the Debye length, which depends on the concentration of cations in solution, to detect DNA hybridization based on the intrinsic negative charge of DNA. The Debye length is longer in buffer solution with a lower concentration of NaCl and the intrinsic negative charge of DNA is more effective on the channel surface in longer Debye length solution. The shifts in the gate voltage by DNA hybridization with complementary target DNA are 21 mV in 1 mM NaCl buffer solution, 7.2 mV in 10 mM NaCl buffer solution, and 5.1 mV in 100 mM NaCl buffer solution. The sensitivity of FETs to detect DNA hybridization based on charge detection without labeling depends on the Debye length.

Mitochondrial DNA polymorphism in Fourteen Geographical Strains of Drosophila melanogoater (세계 14지역 계통에 대한 초파리 미토콘드리아 DNA의 다형현상)

  • 김봉기
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.218-224
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    • 1988
  • Restriction endonucleases were used to search for intraspecific variation at 32 cleavage sites in mitochondrial DNA(mtDNA) purified from fourteen strains of Drosophila melanogaster helonging to different localities of the world. mtDNA of D. melanogaster was displayed site variation(Hpall, Haelll and Seal endonucleases) and length variation(maxirnum 550bp). Six genotypes, Ml, M2, M3, M4, M6 and M7, could be distinguished based on ihe site types witti a low average of intraspecific substitution rate (1.88%),but M5 type of Ogasawara strain in Japan was not detected in this study. A possible explanation for the low divergence was that mtDNA variation of fourteen strains in D. melanogaster could not he accumulated sufficiently owing to recent divergence of few individuals, and that sequence divergence was prevented by frequent migration in spite of the geographical isolation.

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Changes in Polyamine Level and Chloroplast DNA Methylation in Chlamydomonas reinhardtii (Chlamydomonas의 Polyamine 함량변화와 엽록체 DNA Methylation)

  • 이순희
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.101-109
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    • 1994
  • Relationship between polyamine level and DNA methylation in the absence or presence of MGBG(l mM), which is an enzyme-activated reversible inhibitor of SAMDC, has been investigated during gametogenesis of Chlamydomonas. In the absence of MGBG, polyamine levels decreased in Chlamydomonas 137C(+) and 137C(-) during gametogenesis. And polyamine level of 137C(+) was 2-5 times as much as that of 137C(-) and showed a significant decrease unlike that of 137C(-). In vitro, MGBG inhibited ctDNA methylation of 137C(+) by 20-30% but did not inhibited that of 137C(-). Also, MGBG inhibited DNA methylase by 60% in vitro. The results obtained in the present work suggest the possibility that the changes of polyamine level may be associated with ctDNA methylation during gametogenesis of Chlamydomonas.omonas.

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Detection of Mycoplasmas DNA in the Cancer and the Normal Tissues from the Patients with Gastric and Colon Cancer (위암 및 결장암 조직과 그 주변의 정상조직에서 Mycoplasmas DNA의 정색)

  • Chang, Myung-Woong;Shin, Hyun-Chul;Park, In-Dal;Kim, Kwang-Hyuk
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.279-285
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    • 2007
  • Detection of Mycoplasma DNA from the 30 cases of cancer tissues and the normal tissues surrounding the cancer tissues obtained from the patients with gastric cancer and the other 30 cases of cancer tissues and the normal tissues surrounding the cancer tissues obtained from the patients with colon cancer were evaluated by polymerase chain reaction(PCR). The PCR products were sequenced using an ABI 377 automatic DNA sequencer, and these sequences were confirmed by comparing sequences with the database of the National Center for Biotechnology Information BLAST network server. Mycoplasmas DNA were defected in 18 (60%) cases of normal tissues which were around gastric cancer and were 13 (43.3%) cases of gastric cancer tissues. Mycoplasmas DNA were detected in 15(50%) cases of normal tissues which were around colon cancer and 12 (40%) cases of colon cancer tissues. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, and M. conjunctivae were detected from the gastric cancer tissues. The M. faucium, M. subdolum,, M. salivarium, M. auris, M. hyosynoviae, M. bovigenitalium and M. pulmonis were detected from the normal tissues around gastric cancer. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. synoviae M. bovigenitalium, M. gallinarum, and M. moatsii were detected from the colon cancer. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. bovis, M. opalescens, M. bovigenitalium, M. gallinarum, and M. moatsii were detected from the normal tissues around the colon cancer. These results suggest that Mycoplasmas infection may not correlate with gastric cancer and colon cancer, because of the detection rate of Mycoplasmas DNA were not significantly differences between normal and cancer tissues from the patients.

CpG-DNA induces bacteria-reactive IgM enhancing phagocytic activity against Staphylococcus aureus infection

  • Kim, Te Ha;Kim, Dongbum;Lee, Heesu;Kwak, Min Hyung;Park, Sangkyu;Lee, Younghee;Kwon, Hyung-Joo
    • BMB Reports
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    • v.52 no.11
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    • pp.635-640
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    • 2019
  • CpG-DNA triggers the proliferation and differentiation of B cells which results in the increased production of antibodies. The presence of bacteria-reactive IgM in normal serum was reported; however, the relevance of CpG-DNA with the production of bacteria-reactive IgM has not been investigated. Here, we proved the function of CpG-DNA for the production of bacteria-reactive IgM. CpG-DNA administration led to increased production of bacteria-reactive IgM both in the peritoneal fluid and serum through TLR9 signaling pathway. When we stimulated B cells with CpG-DNA, production of bacteria-reactive IgM was reproduced in vitro. We established a bacteria-reactive monoclonal IgM antibody using CpG-DNA stimulated-peritoneal B cells. The monoclonal IgM antibody enhanced the phagocytic activity of RAW 264.7 cells against S. aureus MW2 infection. Therefore, we suggest that CpG-DNA enhances the antibacterial activity of the immune system by triggering the production of bacteria-reactive IgM. We also suggest the possible application of the antibodies for the treatment of antibiotics-resistant bacterial infections.

Effect of Polyamines on purified DNA Methyltransferase from Chlamydomonas reinhardtii (Chlamydomonas reinhardtii로부터 분리, 정제된 DNA Methyltransferase 활성에 대한 Polyamine의 영향)

  • 이명민
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.331-341
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    • 1989
  • DNA methyltransferase was purified 282.6-fold from Chlamydomonas reinhardtii 21gr (mt+) gametic cell to examine the effect of polyamine on the enzyme acctivity. Polyacrylamide gel electrophoresis(PAGE) revealed at least three bands(1 major band, 2 minor bands). Among these, the major band represents DNA methyltransferase. Polyacrylamide gel electrophoresis in the presence of 0.1% sodium dodecylsulfate(SDS-PAGE) revealed a major band with M.W. 60,000. DNA methyltransferase activity was inhibited more effectively by spermine than by spermidine, and the inhibition by putrescine was smaller than spermine and spermidine. DNA methyltransferase activity was inhibited by 40% and 53% at 5mM and 20mM spermine, respectively. In the case of spermidine, the inhibition was 35% at 10mM and 44% at 20mM. However, the inhibition by putrescine appeared only above 5mM and reached about 25% at 20mM.

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Effect of Low-Energy Electron Irradiation on DNA Damage by Cu2+ Ion

  • Noh, Hyung-Ah;Park, Yeunsoo;Cho, Hyuck
    • Journal of Radiation Protection and Research
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    • v.42 no.1
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    • pp.63-68
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    • 2017
  • Background: The combined effect of the low energy electron (LEE) irradiation and $Cu^{2+}$ ion on DNA damage was investigated. Materials and Methods: Lyophilized pBR322 plasmid DNA films with various concentrations (1-15 mM) of $Cu^{2+}$ ion were independently irradiated by monochromatic LEEs with 5 eV. The types of DNA damage, single strand break (SSB) and double strand break (DSB), were separated and quantified by gel electrophoresis. Results and Discussion: Without electron irradiation, DNA damage was slightly increased with increasing Cu ion concentration via Fenton reaction. LEE-induced DNA damage, with no Cu ion, was only 6.6% via dissociative electron attachment (DEA) process. However, DNA damage was significantly increased through the combined effect of LEE-irradiation and Cu ion, except around 9 mM Cu ion. The possible pathways of DNA damage for each of these different cases were suggested. Conclusion: The combined effect of LEE-irradiation and Cu ion is likely to cause increasing dissociation after elevated transient negative ion state, resulting in the enhanced DNA damage. For the decrease of DNA damage at around 9-mM Cu ion, it is assumed to be related to the structural stabilization due to DNA inter- and intra-crosslinks via Cu ion.

Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai

  • Lee, Soo-Ung;Huh, Sun;Sohn, Woon-Mok;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.129-135
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    • 2004
  • We compared the DNA sequences of the genus Metagonimus: M. yokogawai, M. takahashii, and M. miyatai. We obtained 288 D1 ribosomal DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (mtCOI) fragments from the adult worms by PCR, that were cloned and sequenced. Phylogenetic relationships inferred from the nucleotide sequences of the 28S D1 rDNA and mtCOI gene. M. takahashii and M. yokogawai are placed in the same clade supported by DNA sequence and phylogenie tree analysis in 28S D1 rDNA and mtCOI gene region. The above findings tell us that M. takahashii is closer to M. yokogawai than to M. miyatai genetically. This phylogenetic data also support the nomination of M. miyatai as a separate species.

Bacterial Virus DNA Damage Caused by Fumonisin B1 (Fumonisin B1에 의한 세균바이러스 DNA손상)

  • 이길수;조성국
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.34-38
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    • 1999
  • Fumonisin B1 is a secondary metabolite of Fusarium moniliforme, a contaminant of corn and corn product. Fumonisin B1 has been shown to be responsible for major toxicological effects of the fungus in rats, horses, and pigs. Fumonisin B1 induced λ DNA fragmentation, which was increased with incubation time, reducing agent NADPH and metal ion (Cu2+). The DNA damage was inhibited by dimethyl sulfoxide (DMSO) or mannitol as radical scavenger for free radicals. DNA fragmentation, induced by fumonisin B1 in the presence of 1 mM NADPH and 0.1 mM CuCl2, was inhibited by 100 mM DMSO. By the in vitro reaction of fumonisin B1 with supercoiled plasmid pBR322 DNA, plasmid DNA was relaxed, eventually linearized in the agarose gel electrphoresis. From rifampicin sensitive E. coli CSH138 in bacterial mutagenesis system, the rifampicin resistant E. coli mutants were obtained by fumonisin B1. These results suggest that fumonisin B1 may be a possible environmental mutagen in bacterial mutagen assay system.