• Title/Summary/Keyword: DNA Synthesis

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Effect of Polyacetylene Compounds from Panax Ginseng on Macromolecule Synthesis of Lymphoid lukemia L1210 (인삼 포리아세틸렌 화합물이 Lymphoid lukemia L1210의 고분자물질 합성에 미치는 영향)

  • Kim, Young-Sook;Kim, Shin-Il;Hahn, Dug-Ryong
    • YAKHAK HOEJI
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    • v.32 no.2
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    • pp.137-140
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    • 1988
  • To investigate polyacetylene compounds isolated from petroleum ether extract of panax ginseng effect on the macromolecule synthesis, lympoid lukemia L1210 cell was incubated with them at 4, 8, 12,16 hours. Panaxydol, panaxynol and panaxytriol as cytotoxic substances inhibited the synthesis of macromolecule such as DNA, RNA and protein. Panaxydol which had the most potent cytotoxicity among these three compounds showed the strongest inhibitory effect on DNA, RNA and protein synthesis. For DNA and RNA synthesis, panaxynol and panaxytriol decreased the rate of inhibition with the incubation time but panaxydol had a strongest inhibitory effect at 16 hour incubation time. Protein synthesis was markedly inhibited by all these polyacetylene compounds. It was obserbed that there is a relationship between cytotoxicities of polyacetylene compounds and the inhibition of macromolecule synthesis.

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Preparation of Oligonucleotide Arrays with High-Density DNA Deposition and High Hybridization Efficiency

  • Park, Jeong-Won;Jung, Yong-Won;Jung, Young-Hwan;Seo, Jeong-Sun;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.25 no.11
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    • pp.1667-1670
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    • 2004
  • In DNA microarray produced by DNA-deposition technology, DNA-immobilization and -hybridization yields on a solid support are most important factors for its accuracy and sensitivity. We have developed a dendrimeric support using silylated aldehyde slides and polyamidoamine (PAMAM) dendrimers. An oligonucleotide array was prepared through a crosslinking between the dendrimeric support and an oligonucleotide. Both DNAimmobilization and -hybridization yields on the solid support increased by the modification with the dendrimers. The increase of the immobilization and hybridization efficiency seems to result from a threedimensional arrangement of the attached oligonucleotide. Therefore, our dendrimeric support may provide a simple and efficient solution to the preparation of DNA microarrays with high-density DNA-deposition and high hybridization efficiency.

Chronic Treatment of Ethanol Inhibits Proliferation of Normal Fibroblasts, but Not Oncogenic ras-Transformed Cells

  • Gu, Young-Hwa;Park, Mi-Sun;Jhun, Byung-H.
    • Biomolecules & Therapeutics
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    • v.6 no.4
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    • pp.345-350
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    • 1998
  • The adverse effects of ethanol on cell proliferation have been described for a variety of tissues and cells. In the present study, we investigated whether chronic ethanol intoxication impairs the cell proliferation and DNA synthesis induced by oncogenic $H-ras^{V12}$ - and $v-K-ras^{V12}$-transformed cells. Ethanol treatment inhibited the cell proliferation and the DNA synthesis of control parental fibroblasts in a time- and dose-dependent manner. In contrast, ethanol did not suppress the proliferation of either oncogenic $H-ras^{V12}$ - or $v-K-ras^{V12}$ -transformed fibroblasts. Microinjection of oncogenic $H-Ras^{V12}$ protein induces DNA synthesis and ethanol treatment did not interfere with the DNA synthesis. The antiproliferative toxicity of ethanol was rescued by antioxidants, such as N-acetylcysteine and 4-methlpyrazole. These results indicate that the antiproliferative action site of ethanol toxicity lies upstream or is independent of Ras and ethanol exerts its toxicity through a free radical formation.

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Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus (인유두종바이러스 검출을 위한 상용화된 cDNA 합성 키트의 평가)

  • Yu, Kwangmin;Park, Sunyoung;Chang, Yunhee;Hwang, Dasom;Kim, Geehyuk;Kim, Jungho;Kim, Sunghyun;Kim, Eun-Joong;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.309-315
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    • 2019
  • Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the $R^2$ and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P<0.001 in the $10^2$ and $10^3$ SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the $10^1$ and $10^2$ HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P<0.01 in 8, 4, and 1 ml and P<0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.

Enviromental Toxic Agents on Genetic Material and Cellular Activity III. DNA Polymerase Inhibitors on Repair of Mutagen-Induced DNA Damage in Mammalian Cells (환경성 유해요인이 유전물질과 세포활성에 미치는 영향 III. 포유동물세포에서 돌연변이원에 의한 DNA 상해의 회복에 미치는 DNA 중합효소저해제의 영향)

  • 엄경일;선우양일;이천복;신은주
    • Environmental Mutagens and Carcinogens
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    • v.8 no.1
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    • pp.1-12
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    • 1988
  • The effects of aphidicolin (APC), an inhibitor of DNA polymerase alpha, or 2', 3'-dideoxythymidine 5'-triphosphate (ddTTP), an inhibitor of DNA polymerase beta, on the repair of DNA damage induced by ethyl methanesulfonate (EMS) or bleomycin (BLM) were investigated in Chinese hamster ovary (CHO)-K1 cells. Three assays were employed in this study: unscheduled DNA synthesis, alkaline elution and alkaline sucrose gradient sedimentation. It was shown that APC or ddTTP inhibited DNA induced by EMS, and thus, the post-treatment with APC or ddTTP following EMS treatment was resulted in the more amount of unscheduled DNA synthesis, and the more accumulation of DNA single-stand breaks than the cells post-incubated without APC or ddTTP. While, in the BLM induced DNA repair, only ddTTP inhibited DNA repair induced by BLM. And thus, the groups post-incubated with or without APC after BLM treatment had the same value in the amount of unscheduled DNA synthesis and of DNA single-strand breaks, while post-treatment with ddTTP was resulted in the increased amount of unscheduled DNA synthesis and the increased DNA sin -strand breaks than the group without ddTTP. These results suggested that both of DNA polymerase $\alpha$ and $\beta$ participated in the repair of DNA damage induced by EMS, but in BLM-induced DNA repair, polymerase $\beta$ participated.ipated.

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Synthesis of 1-$\beta$-D-Arabinofuranosylcytosine-5'-methylthioacetate and Evaluation of Its Inhibitory Effect on DNA Synthesis (1-$\beta$-D-Arabinofuranosylcytosine-5'-methylthioacetate의 합성 및 이의 DNA 합성애 대한 억제작용 평가)

  • 이희주;송민경
    • YAKHAK HOEJI
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    • v.30 no.5
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    • pp.238-244
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    • 1986
  • As one of the starting materials, methylthioacetyl chloride(7) was synthesized in fair yield from mercaptoacetic acid via methyl methylthioacetate(5) prepared by alkylation employing N, N'-dicyclohexyl-O-methylisourea(4). Then 1-$\beta$-D-arabinofuranosylcytosine-5'-methylthioacetate (3) was prepared by esterification of ara-C with obtained methylthioacetyl chloride and tested for inhibitory activity on DNA synthesis in the growing primary hepatocytes and hepatoma strains($H_4$-II-E and HTC cells). In these in vitro cell lines, the inhibitory effect of ara-C-MTA(3) on DNA synthesis was similar to that of its parent ara-C but slightly lower.

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Antitumor Activity of Crude Sesaminol in Sesame Seed

  • Ryu, Su-Noh;Lee, Bong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.3
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    • pp.168-171
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    • 1998
  • Sesaminol in sesame seed was postulated to have antitumor activity. The present study was performed to characterize the role of crude sesaminol extracted from sesame seed (Sesame Crude Sesaminol; SCS) on inhibiting the in vitro growth of human leukemia HL-60 cells. SCS inhibited the growth of human leukemia HL 60 cells in culture and macromolecular synthesis in a dose and time dependent manner. The cytostatic range of SCS concentration was found to be 60 to 100 $\mu\textrm{g}$/ml. SCS concentration greater than 200 $\mu\textrm{g}$/mlwere cytocidal to HL-60 cells. When SCS concentraction was 6 $\mu\textrm{g}$/mland 50 $\mu\textrm{g}$/ml the synthesis of HL-60 cells was inhibited by 35% for DNA, 6% for RNA and 5% for protein and 83% for DNA, 76% for RNA and 60% for protein. Of specific interest was the irreversible effect of SCS in inhibiting DNA synthesis of HL-60 cells. This was evidenced from the fact that, even after washed with PBS three times, preincubated HL-60 cells still showed the inhibited DNA synthesis.

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Effects of Ultraviolet Light on DNA Replication and Repair in Cultured Myoblast Cells of Chick Embryo (培養한 鷄胚筋細胞의 DNA複製 및 回復에 미치는 紫外線의 影響)

  • Park, Sang-Dai;Lee, Suck-Hwe;Choe, Soo-Young;Ha, Doo-Bong
    • The Korean Journal of Zoology
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    • v.25 no.2
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    • pp.55-62
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    • 1982
  • DNA synthesis, unscheduled DNA synthesis, excision of pyrimidine dimers and phtoreactivation were determined in UV-irradiated differentiating muscle cells at various times of primary culture of 12 day chick embryos and results obtained were as follows. The rates of UV-induced unscheduled DNA synthesis were increased as increase of UV dose. And the rates were gradually decreased as the increase of time after culture, but at higher doses the decreasing tendency was remarkable. The patterns of DNA replication were changed drastically as a function of time so that in the seven day cultures the rate of $^3$H-thymidine incorporation was found to be 0.2% of the original activity. The pattern of inhibition of DNA replication by UV damage demonstrated that in cells of earlier stages there were no remarkable changes, but in cells of later stages there was significant fluctuation. Photoreactivation and the excision of pyrimidine dimer in the one day cultures showed that photoreactivation occurred immediately after UV-irradiation, but excision of pyrimidine dimer was gradually and slowly occurred. These results indicate that the differentiation of embryonic muscle cells accompanies the gradual reduction of DNA replication and unscheduled DNA synthesis, and that the photoreactivation is rapid process compared to excision repair.

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THE EFFECT OF SODIUM FLUORIDE AND SODIUM ORTHOVANADATE ON OSTEOBLASTIC CELL LINE MC3T3-E1 CELLS (Sodium fluoride와 Sodium orthovanadate가 조골세포주 MC3T3-E1에 미치는 영향에 관한 연구)

  • Kim, Won-Jin;Chung, Kyu-Rhim
    • The korean journal of orthodontics
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    • v.21 no.1 s.33
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    • pp.97-111
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    • 1991
  • It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.

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Biochemical and Molecular Biological Studies on the DNA Replication of Bacteriophage T7 (Bacteriophage T7의 유전자 복제기작에 관한 생화학적, 분자생물학적 특성 연구)

  • KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.2
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    • pp.209-218
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    • 1995
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

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