• The Korean Journal of Zoology
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• v.20 no.1
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• pp.41-48
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• 1977

Dose response of DNA repair synthesis induced by bleomycin was dose-dependent in lower doses, and maximum rate of it at 5 $\\mu$g/ml represents about 15% of total cells analyzed. At higher doses DNA-repair synthesis was reduced and the rate of it remained unchanged even prolonged treatment. Pretreatment with BUdR or IUdR was found to enhance DNA repair synthesis and also to interfere with semiconservative DNA synthesis at higher doses. Time dependence study showed that DNA repair synthesis occurred as long as for 24 hours after removal of bleomycin. These results seem to suggest that bleomycin is not to be an effective chemical in inducing excision repair and that damages induced in DNA by this drug might include not only strand breaks but other types of DNA damage.

• Korean Journal of Agricultural Science
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• v.30 no.1
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• pp.76-88
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• 2003

This study exploits the robot system which is necessary in gene study, bio-technology industry. As well, it can achieve the job of DNA chip manufacturing whose use rate has been increased recently. The robot consists of DNA spotting device for spotting DNA on the silylated slide and well plate, bed for fixing well-plate, washing & drying device of washing and drying the pin part of DNA spotting device, distillation-water vessel, and discharge vessel of wash water. We made the term of sticking DNA to the pin on well plate to be 15 seconds. The spot size of DNA was set to be 0.28 mm on the average by bringing the slide into contact with pin for 1 second. At this rate, if DNA is spotted in the minimum space possible of about 0.32mm, it can stick about 8,100 DNA spots on the well plate. Analyzing the procedure: Movement starts. Pin washes, dries, and smears DNA on the well plate. Spots DNA onto 12 chips takes 2 minutes and 50 seconds.

• Journal of Life Science
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• v.14 no.1
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• pp.131-140
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• 2004

T7 bacteriophage gp4 is the replicative DNA helicase that unwinds double-stranded DNA by utilizing dTTP hydrolysis energy. The quaternary structure of the active form of T7 helicase is a hexameric ring with a central channel. Single-stranded DNA passes through the central channel of the hexameric ring as the helicase translocates $5'\rightarrow3'$ along the single-stranded DNA. The DNA unwinding was measured by rapid kinetic methods and showed a lag before the single-stranded DNA started to accumulate exponentially. This behavior was analyzed by a kinetic stepping model for the unwinding process. The observed lag phase increased as predicted by the model with increasing double-stranded DNA length. Trap DNA added in the reaction had no effect on the amplitudes of double-stranded DNA unwound, indicating that the $\tau7$ helicase is a highly processive helicase. Global fitting of the kinetic data to the stepping model provided a kinetic step size of 10-11 bp/step with a rate of $3.7 s^{-1}$ per step. Both the mechanism of DNA unwinding and dTTP hydrolysis and the coupling between the two are unaffected by temperature from $4∼37^{\circ}C$. Thus, the kinetic stepping for dsDNA unwinding is an inherent property of tile replicative DNA helicase.

• Journal of Radiation Protection and Research
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• v.33 no.2
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• pp.53-59
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• 2008

We observed DNA damages as a function of mean absorbed dose to identify the indirect effect of high-energy radiation such as x-ray. Monolayer films of lyophilized pGEM-3Zf(-) plasmid DNA deposited on tantalum foils were exposed to Al $K{\alpha}$ X-ray (1.5 keV) for 0, 3, 7 and 10 min, respectively, in a condition of ultrahigh vacuum state. We compared DNA damages by X-ray irradiation with those by 3 eV electron irradiation. X-ray photons produced low-energy electrons (mainly below 20 eV) from the tantalum foils and DNA damage was induced chiefly by these electrons. For electron beam irradiation, DNA damage was directly caused by 3 eV electrons. Irradiated DNA was analyzed by agarose gel electrophoresis and quantified by ImagaQuant program. The quantities of remained supercoiled DNA after irradiation were linearly decreased as a function of mean absorbed dose. On the other hand, the yields of nicked circular (single strand break, SSB) and interduplex crosslinked form 1 DNA were linearly increased as a function of mean absorbed dose. From this study, it was confirmed that DNA damage was also induced by low energy electrons ($0{\sim}10\;eV$) even below threshold energies for the ionization of DNA.

• Korean Journal of Heritage: History & Science
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• v.40
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• pp.387-411
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• 2007

The analysis of ancient DNA (aDNA) has become increasingly considerable anthropological, archaeological, biological and public interest. Although this approach is complicated by the natural damage and exogenous contamination of a DNA, archaeologists and biologists have attempted to understand issues such as human evolutionary history, migration and social organization, funeral custom and disease, and even evolutionary phylogeny of extinct animals. Polymerase chain reaction(PCR) is powerful technique that analyzes DNA sequences from a little extract of an ancient specimen. However, deamination and fragmentation are common molecular damages of aDNA and cause enzymatic inhibition in PCR for DNA amplification. Besides, the deamination of a cytosine residue yielded an uracil residue in the ancient template, and results in the misincorporation of an adenine residue in PCR. This promotes a consistent substitution (cytosine thymine, guanine adenine) to original nucleotide sequences. Contamination with exogenous DNA is a major problem in aDNA analysis, and causes oversight as erroneous conclusion. This report represents serious problems that DNA modification and contamination are the main issues in result validation of aDNA analysis. Now, we introduce several criterions suggested to authenticate reliance of aDNA analysis by many researchers in this field.

• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.89-91
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• 1998

Cryptotanshinone induced topoisomerase I-mediated DNA cleavage in vitro as strongly as camptothecin, whereas topoisomerase II-mediated DNA cleavage was not induced by this agent. In DNA relaxation assay using calf thymus DNA topoisomerase I and supercoiled pBR322 DNA, cryptotanshinone inhibited topoisomerase I-mediated DNA relaxation in a dose-dependent manner. In unwinding assay, cryptotanshinone ($50{\mu}M$) did not shift the topoisomers of DNA. These results suggest that cryptotanshinone exerted a preferential inhibition of topoisomerase I without intercalating into DNA.

• Development and Reproduction
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• v.2 no.2
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• pp.197-203
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• 1998

DNA methylation seems to play an important regulatory role in gene expression and cell differentiation during postimplantation embryonic development. However, the significance of DNA methylation which is maintained by the DNA MTase during preimplantation embryonic development, is not fully understood. In order to study the role of DNA methylation in the preimplantation embryos, the expression of DNA MTase transcripts was monitored in the oocytes and preimplantation embryos. The mRNA of DNA MTase was detected in the oocytes and pleimplantation embryos. The relative mRNA levels of DNA MTase were high from the stages of GV-oocytes and pronuclear embryos, and thereafter decreased gradually. By the treatment of $\alpha$-amanitin, it was confirmed that the transcripts presented in pronuclear embryos was derived from the maternal genome. The presence of transcripts of DNA MTase in the oocytes and pronuclear embryos suggests that the maintenance of DNA methylation may be necessary and seems to play an important role in gene expression and cell differentiation during preimplantation embryonic develop-ment in mouse.

• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.253-258
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• 2004

DNA double-strand break (DSB) is a serious treat for the cells including mutations, chromosome rearrangements, and even cell death if not repaired or misrepaired. Ku heterodimer regulatory DNA binding subunits (Ku70/Ku80) bound to double strand DNA breaks are able to interact with 470-kDa DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and the interaction is essential for DNA-dependent protein kinase (DNA-PK) activity. The Ku80 mutants were designed to bind Ku70 but not DNA end binding activity and the peptides were treated in breast cancer cells for co-therapy strategy to see whether the targeted inhibition of DNA-dependent protein kinase (DNA-PK) activity sensitized breast cancer cells to ionizing irradiation or chemotherapy drug to develop a treatment of breast tumors by targeting proteins involved in damage-signaling pathway and/or DNA repair. We designed domains of Ku80 mutants, 26 residues of amino acids (HN-26) as a control peptide or 38 (HNI-38) residues of amino acids which contain domains of the membrane-translocation hydrophobic signal sequence and the nuclear localization sequence, but HNI-38 has additional twelve residues of peptide inhibitor region. We observed that the synthesized peptide (HNI-38) prevented DNA-PKcs from binding to Ku70/Ku80, resulting in inactivation of DNA-PK complex activity in breast cancer cells (MDA-465 and MDA-468). Consequently, the peptide treated cells exhibited poor to no DNA repair, and became highly sensitive to irradiation or chemotherapy drugs. The growth of breast cancer cells was also inhibited. These results demonstrate the possibility of synthetic peptide to apply breast cancer therapy to induce apoptosis of cancer cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.6
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• pp.569-572
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• 1987

The DNA damage by linoleic acid hydroperoxide (LHPO) was investigated in a DNA-LHPO system at $37^{\circ}C$ to elucidate the DNA damage mechanism by lipid peroxidation products. LHPO shelved a great DNA damage with the increase of its concentrations. DNA was completely damaged in a LHPO-DNA(weight ratio, 2:3) system after incubation for 2 days. The degree of DNA ,damage by LHPO was greated than that of linoleic acid. In the quantitative analysis of DNA damage, the decreasing ratio of DNA content was $60\%$ in $84{\mu}g$ LHPO system incubated for 1 day compared to the control solution marked $30\%$. There were no participation of active oxygens on the DNA damage by LHPO.

• Polymer(Korea)
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• v.29 no.1
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• pp.69-73
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• 2005

To overcome the main disadvantages of non-viral gene delivery systems such as repeated administration due to the low transfection efficiency, poly(D,L-lactide-co-glycolide) was applied to encapsulate pDNA in its microsphere formulation. Free pDNA or various ratios (w/w) of chitosan/pDNA complexes was used for encapsulation, with the resulting encapsulation efficiency of 44%, 5%, and 8% for free pDNA, 0.7:1 and 1:1 ratios, respectively. Scanning electron micrographs of poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres encapsulating pDNA or chitosan-condensed pDNA revealed a smooth spherical shape immediately after microsphere preparation and a collapsed porous shape in 41 days due to the degradation of PLGA. In vitro release profile showed that the 0.7:1 (w/w) ratio formulation exerted 47% release in 26 days, whereas free pDNA or 1:1 (w/w) ratio formulation did only 15% or 32%, respectively.