• Title/Summary/Keyword: DnaB

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Diverse Mutations of rpoB in Rifampin-Resistant Mycobacteria (Rifampin에 대한 내성 마이코박테리아에서 rpoB의 다양한 변이)

  • Kweon, Tae-Dong;Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.10a
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    • pp.991-993
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    • 2012
  • We analyzed RNA polymerase beta subunit gene (rpoB) mutation of rifampin-resistant Mycobacteria through analysis of nucleotide sequence of rpoB DNA (351 bp) containing rifampin resistant region, $rif^r$. For this study, we collected rifampin-resistant Mycobacteria that were identified by conventional culture methods from Masan National Hospital and The Korean Institute of Tuberculosis. We performed sequencing of DNA nucleotides and analyzed rpoB gene of those rifampin-resistant Mycobacteria. From this analysis, we invcestigated diverse mutations of rpoB gene included rifampin-resistant gene, which were not reported, from those rifampin-resistant Mycobacteria.

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Antigenotoxicity of Ginseng Petroleum Ether Extract and its Action Mechanism (인삼 지용성성분인 유전독성억제효과와 작용기전)

  • 허문영
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.243-251
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    • 1998
  • Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatic agent. Petroleum ether extract of panax ginseng C.A. Meyer (GPE) and its several fractions (PI-P5) were tested for the evaluation of antigenotoxicity against N-methyl-N-nitrosourea (MNU) and benzo(a)pyrene [B(a)P]-induced micronucleated reticulocytes in mouse peripheral blood. GPE and P2 showed more significant anticlastogenicity than other fractions did. To elucidate the anticlastogenic action mechanism of GPE and P2 against B(a)P, the alteration of B(a)P metabolism was studied. GPE and P2 inhibited B(a)P metabolism in the presence of 8-9 mix and decreased B(a)P-DNA binding in calf thymus DNA with 8-9 mix. They also decreased [$^3H$] MNU induced DNA binding and methylation to 7-methyl guanine and $O^{6}-methyl$ guanine adducts in calf thymus DNA by RPLC analysis. These results suggest that the anticlastogenicity of GPE and P2 on the B(a)P or MNU-induced clastogenicity is due to decrease of DNA binding with B(a)P or MNU, the inhibition of metabolism with B(a)P and the inhibition of methylation in DNA. Therefore, GPE and P2 may be useful chemopreventive agents of alkylating agent like MNU and secondary carcinogen like B(a)P.

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ROLE OF NF${\kappa}B$ IN TOLL-LIKE RECEPTOR 9-MEDIATED MATRIX METALLOPROTEINASE-9 EXPRESSION (Toll-like receptor 9-매개에 의한 matrix metalloproteinase-9 발현에서 NF${\kappa}B$의 역할)

  • Lee, Sang-Hoon;Chin, Byung-Rho;Baek, Suk-Hwan
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.6
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    • pp.636-642
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    • 2007
  • Background: CpG DNA plays an important role in immune cell function. This study examined whether the temporal control of toll-like receptor (TLR)9 by CpG DNA can regulate the expression of matrix metalloproteinase-9(MMP-9). Methods and materials: Macrophages were cultured in the presence of 10% FBS. For the various MMP genes analysis, RT-PCR and real-time PCR were performed. In addition, zymography assay performed for the MMP activity. The phosphorylation assay did for the ERK1/2 and NF${\kappa}B$ activation, and luciferase promoter assay was for the NF${\kappa}B$ activity. Results: CpG DNA induced the mRNA expression of MMP-2, MMP-9, and MMP-13, but not of MMP-7, MMP-8, and MMP-12, in a time-dependent manner. Especially, the mRNA expression of MMP-9 was strongly induced by CpG DNA using real-time RT-PCR. The TLR9 inhibitor, chloroquine, suppressed CpG DNA-induced MMP-9 expression and its activity. Moreover, CpG DNA induced the phosphorylation of ERK and the inhibition of ERK by U0126 suppressed CpG DNA-induced MMP-9 expression and its activity. CpG DNA stimulated $I{\kappa}B-{\alpha}$ degradation and luciferase activity. In addition, pretreatment of SN-50, the inhibitor of NF${\kappa}B$, strongly blocked the CpG DNA-induced MMP-9 expression and activity. Conclusion: These observations suggest that CpG DNA may play important roles in the activation of macrophages by regulating the production of MMP-9 via the sequential TLR9-ERK-NF${\kappa}B$ signaling pathway.

DNA Computing Adopting DNA Coding Method to solve Maximal Clique Problem (Maximal Clique Problem을 해결하기 위한 DNA 코딩 방법을 적용한 DNA 컴퓨팅)

  • Kim, Eun-Kyoung;Lee, Sang-Yong
    • The KIPS Transactions:PartB
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    • v.10B no.7
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    • pp.769-776
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    • 2003
  • DNA computing has used to solve MCP (Maximal Clique Problem). However, when current DNA computing is applied to MCP. it can't efficiently express vertices and edges and it has a problem that can't look for solutions, by misusing wrong restriction enzyme. In this paper we proposed ACO (Algorithm for Code Optimization) that applies DNA coding method to DNA computing to solve MCP's problem. We applied ACO to MCP and as a result ACO could express DNA codes of variable lengths and generate codes without unnecessary vertices than Adleman's DNA computing algorithm could. In addition, compared to Adleman's DNA computing algorithm, ACO could get about four times as many as Adleman's final solutions by reducing search time and biological error rate by 15%.

Animal species identification by co-amplification of hypervariable region 1 (HV1) and cytochrome b in mitochondrial DNA

  • Lim, Si Keun;Park, Ki Won
    • Analytical Science and Technology
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    • v.18 no.3
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    • pp.257-262
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    • 2005
  • Mitochondrial DNA (mt DNA) sequence analysis has been a useful tool for species identification of animals and human individuals. Two hypervariable regions (HV1 and HV2) in control region of mitochondrial genome were analyzed for human individual identification. In case of animal species identification, several genes on mt DNA such as cytochrome b (cytb), RNAs, cytochrome oxidases (CO) were used. In this study, co-amplification of HV1 and cytb was carried out in order to check the contamination of animal DNA and to verify the human DNA. The primer sets used in PCR were H15997/L16236 for HV1 and H14724/L15149 for cytb. PCR products for HV1 and cytb were 239 bp and 425 bp, respectively. The appearance of two bands on agarose gel implied the DNA came from human, however the single band of cytb gene represented the non-human animal DNA.

Effect of Fumonisin B1 on the Bacterial Virus Multiplication (세균 바이러스 증식에 대한 Fumonisin B1의 영향)

  • 이길수
    • Toxicological Research
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    • v.12 no.1
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    • pp.17-20
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    • 1996
  • The effect of Fumonisin B1, a mycotoxin produced by Fusarium moniliforme on bacterial viruses P1 and Lambda, was investigated by the virus plaque assay. Fumonisin B1 inhibited the P1 viral multiplication in the concentration range from $100{\mu}g$/ml to $400{\mu}g$/ml. The inhibition was Fumonisin B1 concentration-dependent. Another bacterial virus Lambda multiplication was also inhibited by lower concentration of Fumonisin B1 ($10{\mu}g$/ml~$50{\mu}g$/ml). This inhibition was dependent on Fumonisin B1 and on virus-Fumonisin B1 reaction time. Sensitivity of bacteriophage Lambda to Fumonisin B1 was higher than that of P1 virus. Lambda vital DNA was treated in vitro with Fumonisin B1 at various concentration. Significant DNA fragmentation by Fumonisin 191 was observed in the agarose gel electrophoresis. Lambda viral DNA was partially digested even in the Fumonisin B1 $10{\mu}g$ and the level of its fragmentation was dependent on Fumonisin B1 amount up to $30{\mu}g$ per assay.

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DNA Breakage by Salvianolic acid B in the Presence of Cu (II) (구리이온(II)이 존재할 때 Salvianolic acid B에 의한 DNA 절단)

  • Lee, Pyeongjae;Moon, Cheol;Choi, Yoon Seon;Son, Hyun Kyu
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.205-210
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    • 2018
  • Salvianolic acid B, which is a compound in the Salvia miltiorrhiza, has diverse biological activities, In particular, the antioxidative effects were reported to be involved in the protection of hepatocytes, neurons, and various cell types. On the other hand, some phenolic compounds, such as ferulic acid, which is regarded as an antioxidant, plays a pro-oxidative role in the specific transitional metal environment, which could explain the anticancer effect. This study examined the pro-oxidative effects of salvianolic acid B in the presence of $Cu^{2+}$. Treatment with both salvianolic acid B and $Cu^{2+}$ induced the transition of supercoiled DNA to the open circular or linear form but not in the sole salvianolic acid B or $Cu^{2+}$ treatments. Salvianolic acid B reduced the $Cu^{2+}$ to $Cu^+$ using neocuproine, a $Cu^+$ specific chelator. In addition, catalase, an enzyme that breaks down the $H_2O_2$ to water and molecular oxygen, inhibited the DNA breakage. $H_2O_2$, a reactive oxygen species, has detrimental effects on biological molecules, particularly DNA. Overall, the reduction of $Cu^{2+}$ by salvianolic acid B could lead to the production of $H_2O_2$ followed by DNA breakage. These results suggest that the pro-oxidative effects could be the one of the anti-cancer mechanisms of salvianolic acid B, which remains to be explained.

Domain Expression of ErmSF, MLS (macrolide-lincosamide-streptogramin B) Antibiotic Resistance Factor Protein (MLS (macrolide-lincosamide-streptogramin B) 항생제 내성인자 단백질인 ErmSF의 domain발현)

  • 진형종
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.245-252
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    • 2001
  • Erm proteins, MLS (macrolide-lincosamide-streptogramin B) resistance factor proteins, show high degree of amino acid sequence homology and comprise of a group of structurally homologous N-methyltransferases. On the basis of the recently determined structures of ErmC` and ErmAM, ErmSF was divided into two domains, N-terminal end catalytic domain and C-terminal end substrate binding domain and attempted to overexpress catalytic domain in E. coli using various pET expression systems. Three DNA fragments were used to express the catalytic domain: DNA fragment 1 encoding Met 1 through Glu 186, DNA fragment 2 encoding Arg 60 to Glu 186 and DNA fragment 3 encoding Arg 60 through Arg 240. Among the pET expression vectors used, pET 19b successfully expressed the DNA fragment 3 and pET23b succeeded in expression of DNA fragment 1 and 2. But the overexpressed catalytic domains existed as inclusion body, a insoluble aggregate. To assist the soluble expression of ErmSF catalytic domains, Coexpression of chaperone GroESL or Thioredoxin and lowering the incubation temperature to $22^{\circ}C$ were attempted, as did in the soluble expression of the whole ErmSF protein. Both strategies did not seem to be helpful. Solubilization with guanidine-HCl and renaturation with gradual removal of denaturant and partial digestion of overexpressed whole ErmSF protein (expressed to the level of 126 mg/ι culture as a soluble protein) with proteinase K, nonspecific proteinase are under way.

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Structure-function analysis of PRDI DNA polymerase

  • Jung, Guhung
    • The Microorganisms and Industry
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    • v.16 no.3
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    • pp.6-14
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    • 1990
  • PRDI DNA polymerase is the smallest member of the family B DNA polymerase (Jung et al., 1987). This DNA polyerase is specified by bacteriophage PRDI which infects a wide variety of gram-negative bacteria(Mindich and Bamford, 1988). Because PRDI is highly amenable to genetic and biochemical manipulation, it is a convenient model system with which to study structure-function relationships of DNA polymerase molecules. To determine the functional roles of the highly conserved regions of the family B DNA polymerases, we have initiated site-directed mutagenesis with PRD1 DNA polymerase, and our results show that mutations at the conserved regions within PRD1 DNA polymerase inactivate polymerase complementing activity and catalytic activity.

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Molecular Cloning of Chicken Major Histocompatibility Complex Class II Molecules

  • Sung, Aree-Moon
    • Toxicological Research
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    • v.8 no.2
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    • pp.331-342
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    • 1992
  • The chicken major histocompatibility complex (MHC), the B complex, is beginning to be analyzed at the DNA level. Inbred lines of chickens have been reported to possess 3~5 MHC class II genes. To further analyzed the molecular structure of the chicken MHC class II genes, cDNA clones coding for chicken MHC class II (B-L) ${\beta}$ chain molecules were isolated from chicken spleen and liver. Tissue-specific transcription of B-L ${\beta}$genes was studied by Northern blot analysis. A high level of expression was detected for spleen poly(A)$^+$ RNA whereas a faint signal was detected for liver poly(A)$^+$ RNA. Twenty-nine cDNA clones were isolated from the spleen and eight cDNA clones were isolated from the liver. Based on restriction maps, most clones could be clustered into one family of genes. Four cDNA clones were sequenced (S7, S10 and S19 from the spleen and L1, which was identical to S19, from the liver). Complete amino acid sequences of B-L ${\beta}$ chain molecules were predicated from the nucleotide sequences of the cDNA clones. Although both the nature and the location of the conserved residues were similar in chicken and mammalian sequences, some species-specific differences were found, suggesting that the structures of the B-L molecules are similar, but not identical to their mammalian counterparts.

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