• Title/Summary/Keyword: DNA Synthesis

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Analysis of Expressed Sequence Tags Generated from the Posterior Silkgland cDNA Clones of Antheraea yamamai (천잠 후부 견사선 유래 발현 유전자 꼬리표 작성 및 분석)

  • 윤은영;구태원;강석우;이혜원;황재삼;김호락
    • Journal of Life Science
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    • v.10 no.2
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    • pp.188-195
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    • 2000
  • In order to understand molecular events during silk synthesis and provide genetic resources for molecular breeding, we had analyzed the cDNA library constructed from the posterior silkgland of Antheraea yamamai and partially sequenced 276 randomly selected genes from the cDNA library. Database comparisons of the expressed sequence tags (ESTs) revealed that 26 non-redundant clones showed a high similarity with previously identified genes. Among them, 17 clones exhibited a homology with previously identified insect genes and 9 clones were identical to genes that were previously identified from other organisms. A functional categorization showed that silk synthesis-defense- or stress-related genes, as well as genes involved in the metabolic pathways and in the transcriptional or translational apparatus are represented. In this report, the clone (AY479) which had high similarity with fibroin from A. pernyi was particularly analyzed in detail. The AY479 clone was carboxyl terminal region of fibroin. The 472 bp cDNA has 123 amino acids that shared 85% homology with the fibroin from A. pernyi and its deduced peptide had unique feature, that is, sites of alanine rich residues.

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Optimal conditions for pigmentation in Bacillus licheniformis SSA3 and cloning of a DNA fragment involved in pigment production

  • Kim, Jong-Kyu;Shin, Ok-Sun
    • Journal of Microbiology and Biotechnology
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    • v.5 no.1
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    • pp.22-25
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    • 1995
  • Bacillus licheniformis SSA3 can produce a dark-brown antimutagenic pigment. The optimal conditions for production of this pigment are reached at 0.1% tyrosine, in pH 6-8, within 7-9 days, at $30^{\circ}C$, and in aerobic condition. We cloned a DNA fragment involved in pigment synthesis from Bacillus licheniformis SSA3 using a mutant strain. The cloned DNA was 7kb in size, which can produce the same pigment even in E. coli.

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A Visualization Tool for Computational Analysis of DNA Methylation Level Using Bisulfite Sequencing Data

  • Tae, Hong-Seok
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.136-137
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    • 2011
  • Methylation of cytosine is a post-synthesis modification that does not affect the primary DNA sequence but greatly influences gene expression level and phenotypes of an organism. As high-throughput sequencing of bisulfite-treated DNA is the most efficient method to identify methylated sites, several tools to map sequencing reads on a reference are available. But tools to visualize and to interpret the methylation level of methylation sites are currently insufficient. Herein, we present a novel tool to visualize the methylation level of CpG sites.

BrdU에 의한 DNA

  • 손우찬;김형진;이영순
    • Toxicological Research
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    • v.7 no.1
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    • pp.83-92
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    • 1991
  • Complexities of testis structure and function are emphasized in morphometrical and genotoxic evaluation by statistical analysis. F-344 rats were treated with azinphos methyl, cyclophosphomide, and dichlorvos. And Brdu was injected with intrapertionially before sacrifice. The existence and degree of DNA damage were measured by Brdu labeling index which represented relative amount of Brdu incorporated in DNA, morphometric change was evaluated by the relative length of tubular diameter in circular seminiferous tubules and the number of spermatogonia per Sertoli cell in stage IX seminiferous tubules.

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DNA Repair Enhancement by Radioprotective Ginseng Protein Fraction (항 방사선 인삼단백분획의 DNA수복능력 증진효과)

  • Kim, Choon-Mi;Choi, Mi-Kyung
    • YAKHAK HOEJI
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    • v.36 no.5
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    • pp.449-454
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    • 1992
  • The effect of radioprotective ginseng protein fraction on DNA repair capacity was determined by measuring the amount of $^{3}H-thymidine$ incorporated into DNA in the process of repair synthesis for UV damaged DNA. CHO-Kl cells were prepared whose semiconservative replication was inhibited by trimethylpsoralen plus near-UV(PUVA) treatment. When the cells were exposed to UV light alone, the DNA repair capacity was increased at first and then decreased as UV dose increased. However, when the ginseng fraction was treated to the cells, the DNA repair capacity was kept increasing regardless of UV dose increment. When the concentration of protein contained in the added fraction was increased gradually, the repair capacity was also increased almost linearly showing dose-response relationship of the effect. These results suggest that the enhancement of DNA repair capacity of the cell can be one of the mechanisms of radioprotection by the ginseng fraction.

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Effects of the Protein Fraction of Panax ginseng on Primary Cultured Chicken Skeletal Muscle Cells (인삼 단백분획물이 일차배양한 계배의 근육세포에 미치는 영향)

  • Park, Mi-Jung;Song, Jin-Ho;Lee, Heun-Pa;Kim, Young-Choong
    • Korean Journal of Pharmacognosy
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    • v.21 no.3
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    • pp.210-216
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    • 1990
  • Effects of the protein fraction of Panax ginseng on chicken embryonic skeletal muscle cells cultured with a decfiient medium were studied. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton(fraction A), between 5,000 and 10,000 dalton(fraction B), between 1,000 and 5,000 dalton(fraction C), between 500 and 1,000 dalton(fraction D). According to the microscopic observation, all four protein fractions at the concentration of $10{\sim}100{\;}{\mu}g/ml$ showed the tendency to stimulate the growth and differentiation of the muscle cells. The activity of acetylcholinesterase in muscle cells was significantly elevated by the protein fraction A at the concentration of $100{\mu}{\;}g/ml$. Protein fractions B,C and D significantly enhanced the synthesis of RNA in the muscle cells. The synthesis of DNA in muscle cells was significantly enhanced by protein fractions A,B and C.

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PCNA Modifications for Regulation of Post-Replication Repair Pathways

  • Lee, Kyoo-young;Myung, Kyungjae
    • Molecules and Cells
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    • v.26 no.1
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    • pp.5-11
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    • 2008
  • Stalled DNA replication forks activate specific DNA repair mechanism called post-replication repair (PRR) pathways that simply bypass DNA damage. The bypassing of DNA damage by PRR prevents prolonged stalling of DNA replication that could result in double strand breaks (DSBs). Proliferating cell nuclear antigen (PCNA) functions to initiate and choose different bypassing pathways of PRR. In yeast, DNA replication forks stalled by DNA damage induces monoubiquitination of PCNA at K164, which is catalyzed by Rad6/Rad18 complex. PCNA monoubiquitination triggers the replacement of replicative polymerase with special translesion synthesis (TLS) polymerases that are able to replicate past DNA lesions. The PCNA interaction motif and/or the ubiquitin binding motif in most TLS polymerases seem to be important for the regulation of TLS. The TLS pathway is usually error-prone because TLS polymerases have low fidelity and no proofreading activity. PCNA can also be further polyubiquitinated by Ubc13/ Mms2/Rad5 complex, which adds an ubiquitin chain onto monoubiquitinated K164 of PCNA. PCNA polyubiquitination directs a different PRR pathway known as error-free damage avoidance, which uses the newly synthesized sister chromatid as a template to bypass DNA damage presumably through template switching mechanism. Mammalian homologues of all of the yeast PRR proteins have been identified, thus PRR is well conserved throughout evolution. Mutations of some PRR genes are associated with a higher risk for cancers in mice and human patients, strongly supporting the importance of PRR as a tumor suppressor pathway.

Effects of Yuldahansotang water extract on Cultured Primary Hippocampal Cell Culture Damaged by Hydrogen Peroxide (열다한소탕(熱多寒少湯) 전탕액(煎湯液)이 Hydrogen Peroxide에 의해 손상(損傷)된 배양(培養) 해마신경세포(海馬神經細胞)에 미치는 영향)

  • Park, Hye-Sun;Kim, Kyung-Yo;Go, Gi-Deok;Kim, Il-Hwan;Lee, Jae-Heung
    • Journal of Sasang Constitutional Medicine
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    • v.14 no.1
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    • pp.79-89
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    • 2002
  • To evaluate the effect of Yuldahansotang(YHT) water extract on cultured hippocampal cell was inhibited by hydrogen peroxide, MTT assay, NR assay, Neurofilament enzymeimmuno assay and DNA synthesis assay were carried out after the cultured hippocampal cells were preincubated with various concentrations of YHT water extract for 3 hours prior to exposure of hydrogen peroxide. The results obtained were as follows: 1. Hydrogen Peroxide decreased the survival rate of the cultured hippocampal cells on NR assay and MIT assay. 2. YHT water extract have efficacy of increasing a amount of neurofilament decreased by hydrogen peroxide in cultured hippocampal cells. 3. YHT water extract have efficacy of increasing DNA synthesis decreased by hydrogen peroxide in cultured hippocampal cells. From above the results, It is concluded that YHT has marked efficacy in preventing for the damages by hydrogen peroxide.

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Phylogenetic Analysis of Pleurotus Species Based on the Nuclear SSU rRNA Sequences (Phylogenetic Analysis of Pleurotus Species Based on the Nuclear SSU rRNA Sequences)

  • Jeong, Jae Hun;Kim, Eun Gyeong;No, Jeong Hye
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.37-37
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    • 1996
  • The internal regions of nuclear small subunit rRNA from 6 plaeurotus species and 5 Pleurotus ostreatus strains were amplified by PCR and sequenced. The DNA sequences of 8 Pleurotus strains (P. ostreatus NFFA2, NFFA4501, NFFA4001, KFFA4001, KFCC11635, P florida, P. florida, P. sajor-cuju, P. pulmonarius, and P. spodoleucus) were idential, but P. cornucopiae differed from them in two bases out of 605 bases. However, p[hylogenetic analysis of the sequences by DNA-distance matrix and UPGMA methods showed that P. ostreatus NFFA2m1 and NFFA2m2, known as mutants of P. ostreatus NFFA2, belonged to anther group of Basidiomycotina, which is close to the genus Auricularia. The difference of the SSU rDNA sequences of P. cornucopiae from other Pleurotus species tested corresponds to the difference of mitochondrial plasmid type present in Pleurotus species as observed by Kim et al. (1993, Korean J. Microbiol. 31, 141-147).ishement of silencing at the HMR/hsp82 locus can occur in G1-arrested cells. Cell cycle arrest at G1 phase was achieved by treatment of early log a cell cultures with .alpha.-factor mating pheromone, which induces G1 arrest. The result suggests that passage through S phase (and therefore DNA replication) is nor required for re-establishing silencer-mediated repression at the HMNRa/HSP82 locus. Finally, to test whether de nono protein synthesis is required for re-establishment of silencer-mediated repression, cells were pretreated with cycloheximide (500 /.mu.g/ml) 120 min. It was apparent that inhibiting protein synthesis delays, but does not prevent, re-establishment of silencer-mediated repression. Altogether, these results indicate that re-establishment of silencer-mediated repression is not dependent on the DNA replication and has no requirement for protein synthesis.

The Role of Cytoskeletal Elements in Shaping Bacterial Cells

  • Cho, Hongbaek
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.307-316
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    • 2015
  • Beginning from the recognition of FtsZ as a bacterial tubulin homolog in the early 1990s, many bacterial cytoskeletal elements have been identified, including homologs to the major eukaryotic cytoskeletal elements (tubulin, actin, and intermediate filament) and the elements unique in prokaryotes (ParA/MinD family and bactofilins). The discovery and functional characterization of the bacterial cytoskeleton have revolutionized our understanding of bacterial cells, revealing their elaborate and dynamic subcellular organization. As in eukaryotic systems, the bacterial cytoskeleton participates in cell division, cell morphogenesis, DNA segregation, and other important cellular processes. However, in accordance with the vast difference between bacterial and eukaryotic cells, many bacterial cytoskeletal proteins play distinct roles from their eukaryotic counterparts; for example, control of cell wall synthesis for cell division and morphogenesis. This review is aimed at providing an overview of the bacterial cytoskeleton, and discussing the roles and assembly dynamics of bacterial cytoskeletal proteins in more detail in relation to their most widely conserved functions, DNA segregation and coordination of cell wall synthesis.