• 한국환경성돌연변이발암원학회지
• /
• 제15권2호
• /
• pp.88-93
• /
• 1995

Effects of hyperthermia on the induction of DNA single strand breaks and replication inhibition were studied in bleomycin-treated CHO-K$_1$ cells by alkaline elution and alkaline sucrose gradient sedimentation. Bleomycin-induced DNA single strand breaks of DNA were dose-and time-dependently increased, and these strand breaks of DNA were gradually rejoined as post-incubation time passed. Treatment with hyperthermia alone did not affect the induction of DNA single strand breaks. However, pre-exposure of cells to hyperthermia followed by bleomycin treatment greatly increased the single strand breaks, and also reduced the rejoining processes of bleomycin-induced DNA single strand breaks. Bleomycin selectively inhibited the replicon initiation. The combined treatment with hyperthermia and bleomycin markedly potentiated the nonspecific inhibition of replication.

• 대한화학회지
• /
• 제65권2호
• /
• pp.121-124
• /
• 2021

RNA molecules which bind to the G-rich sequence in the 5'-UTR RNA which plays an important role in expression of N-ras, were selected. The secondary structures of five selected RNA aptamers including primer sequence were found by the CLC RNA workbench ver. 4.2 program (www.clcbio.com) and investigated with RNA structural probes such as RNase T1 which has specificity for a G in single-stranded region, RNase V1 specific for double strand and nuclease S1 specific for single strand. The generalized secondary structure model was proposed and characterized. It was composed of a central long double strand region flanked by single strand region at both end sides. The double strand region had an internal single-strand region and bulges. The single strand loop in the right side was composed of four or five nucleotides.

• 한국수산과학회지
• /
• 제36권4호
• /
• pp.346-351
• /
• 2003

Single-cell gel electrophoresis (comet assay) was used to detect DNA single strand break in blood cells from several marine fish species. Three fish species were collected from Georgia coastal area. Mummichog, Fundulus heteroclitus showed higher DNA damage than sea bass, Lateolabrax japonicus and trout, Oncorhynchus masou masou under the same experimental conditions. Mummichogs had more alkaline-labile sites on their DNA than other fish species. The comet assay with mummichog blood cells at pH 12.5 showed a dose-response curve with the increasing concentrations of hydrogen peroxide. While the isolated leucocytes showed no increase of DNA damage after in vitro exposure to 2-methyl-1,4-naphthoquinone (MNQ), erythrocytes showed dose-dependent DNA damage. These results indicate that the comet assay can be applied successfully as a bioassay using erythrocyte for environmental monitoring.

• Toxicological Research
• /
• 제2권1호
• /
• pp.1-7
• /
• 1986

Procarcinogen induced DNA single-strand breaks and unschduled DNA synthesis were measured in primary rat hepatocytes culture. For DNA single-strand breaks assay, rat liver DNA was prelabeled by injection 3H-thymidine during the peak of DNA synthesis following partial hepatectomy. Hepatocytes were isolated from the rat 2 weeks after surgery by a collagenase perfusion techinique and maintained as monolayers in serum free medium on collagen-coated culture dishes. DNA sigle-strand breaks were measured by the alkaline elution techinique.

• Radiation Oncology Journal
• /
• 제8권2호
• /
• pp.137-144
• /
• 1990

Filter elution 방법으로 EL 4백서 백혈병 세포 및 C57BL/6백서 유래의 비장 임파구에 대한 Co-60 $\gamma$ 선의 DNA single-strand breaks (SSB) 효과를 정량적으로 측정하였다. 임파구는 [${^3}H$]thymidine을 표지하기 위하여lipopolysaccharide (LPS, 20 $\mug/ml$)를 첨가하여 자극하고 부유 상태의 EL 4 세포 및 임파구를 $0^{\circ}C$에서 0 Gy, 1 Gy, 5 Gy, 10 Gy 또는 15 Gy조사하였으며, elution용액의 pH는 12.1로 하였다. $\gamma$ 선 조사에 따른 single-strand breaks의 수는 방사선 조사량에 따라 증가 하였으며 21 ml elution 양을 기준으로 한 strand scission factor (SSF)는 EL 4 세포에서 $0.01301\pm0.00096\;Gy^{-1}(n=5)$이었고, 임파구는 $0.01097\pm0.00091\;Gy^{-1}(n=5)$를 나타내므로 본 실험에서는 EL 4 세포가 정상 임파구에 비하여 방사선에 의한 DNA SSB가 민감함을 알 수 있었다(p<0.005). 본 연구 결과 DNA strand breaks의 측정법을 이용하여 방사선의 특성 및 생물학적 효과의 파악은 물론 나아가 기존의 방호제 및 새로운 약제의 DNA에 대한 효과를 판별할 수 있을 것이다.

• Journal of Microbiology and Biotechnology
• /
• 제16권12호
• /
• pp.2004-2007
• /
• 2006

An efficient method for the in vitro reassembly of homologous DNA sequences is presented. The proposed method involves obtaining single strands of homologous genes and hybridizing them to obtain partially hybridized heteroduplex DNA; cleaving the single-stranded regions of the heteroduplex DNA using S1 nuclease to generate double-strand DNA fragments; denaturing the double-strand DNA fragments to generate single-strand DNA fragments; conducting a series of polymerase chain reactions (PCR) using the single-strand DNA fragments as internal primers and a mixture of homologous DNA as templates to obtain elongated reassembled DNA; and finally, amplifying the reassembled DNA by a PCR using terminal primers. As a result, DNA reassembly could be achieved between homologous genes with a sequence similarity as low as 78%.

• Journal of Species Research
• /
• 제9권4호
• /
• pp.448-454
• /
• 2020

Nine free-living heterotrophic flagellates were cultured from marine intertidal sediments and freshwater sediments from Korea. These species are described with uninterpreted records based on light microscopy of living cells and reported taxonomically for the first time from Korea. Diagnostics of these species are as follows; Notosolenus hemicircularis: 9-11.8 ㎛ long with flagellar reservoir, ventrally flattened and dorsally convex with hyaline semicircular collar around short anterior neck, and 8 ridges on cell surface. Thecamonas tranhens: 4.5-7.1 ㎛ long, plastic with proboscis comprising an anterior flagellum surrounded by membranous sleeve. Bodomorpha minima: 4.5-7.0 ㎛ long, rigid with small rostrum in anterior end and active anterior flagellum. Cercomonas hiberna: 5.6-10.9 ㎛ long, very plastic with pseudopodia, cytoplasmic strand and 1 or 2 contractile vacuoles. Cercomonas pellucida: 7.5-13 ㎛ long, plastic with pseudopodia, cytoplasmic strand and single contractile vacuole. With nucleus closely connected to basal bodies. Eocercomonas echina: 4.7-6.5 ㎛ long, plastic with pseudopodia, cytoplasmic strand and 1 or 2 contractile vacuoles. Paracercomonas astra: 5.7-7.3 ㎛ long, moderately metabolic with pseudopodia, cytoplasmic strand and 1 or 2 contractile vacuoles. Paracercomonas minima: 5-9 ㎛ long, metabolic with pseudopodia, cytoplasmic strand and single contractile vacuole. Paracercomonas producta: 6.1-9.9 ㎛ long, very metabolic with pseudopodia, long cytoplasmic strand and single contractile vacuole.

• Bulletin of the Korean Chemical Society
• /
• 제31권7호
• /
• pp.2011-2014
• /
• 2010

The fluorescence intensity of a single-stranded oligonucleotide containing a fluorene-labeled deoxyuridine $(U^{Fl})$ unit increases by only 1.5-fold upon formation of its perfectly matched duplex. To increase the fluorescence signal during hybridization, we positioned a quencher strand containing a deoxyguanine (dG) nucleobase, functioning as an internal quencher, opposite to the $U^{Fl}$ unit to reduce the intrinsic fluorescence upon hybridization with a probe. From an investigation of the optimal length of the quencher strand and the effect of the neighboring base sequence, we found that a short strand (five-nucleotide) containing all natural nucleotides and dG as an internal quencher was effective at reducing the intrinsic fluorescence of a linear beacon; it also exhibited high total discrimination factors for the formation of perfectly matched and single base-mismatched duplexes. Such assays that function based on clear changes in fluorescence in response to single-base nucleotide mutations would be useful tools for accelerating diagnoses related to various diseases.

• Molecular & Cellular Toxicology
• /
• 제1권2호
• /
• pp.99-105
• /
• 2005

Perchloroethylene (tetrachloroethylene, PCE), a dry cleaning and degreasing solvent, can enter ground-water through accidental leak or spills. PCE can be degraded to trichloroethylene (TCE), 1, 1-dichloroethylene (DCE) and vinyl chloride (VC) as potential bio-product. These compounds have been reported that they can cause clinical diseases and cytotoxicity. However, only a little genotoxic information of these compounds has been known. In this study, we investigated DNA single strand breaks of PCE, TCE, DCE and VC by single cell gel electrophoresis assay, (comet assay) which is a sensitive, reliable and rapid method for DNA single strand breaks with mouse lymphoma L5178Y cells. From these results, $37.5\;{\mu}g/ml$ of PCE, $189\;{\mu}g/ml$ of TCE and $56.4\;{\mu}g/ml$ of DCE were revealed significant DNA damages in the absence of S-9 metabolic activation system meaning direct-acting mutagen. And in the presence of S-9 metabolic activation system, $41.5\;{\mu}g/ml$ of PCE, $328.7\;{\mu}g/ml$ of TCE and $949\;{\mu}g/ml$ of DCE were induced significant DNA damage. In the case of VC, it was revealed a significant DNA damage in the presence of S-9 metabolic activation system. Therefore, we suggest that chloroethylene compounds (PCE, TCE, DCE and VC) may be induced the DNA damage in a mammalian cell.

• 방사선산업학회지
• /
• 제5권3호
• /
• pp.243-247
• /
• 2011

In this study, a convenient method for the selective immobilization of single strand DNA (ssDNA) on a polymer surface was described. A positively charged polyelectrolyte, poly(diallyldimethylammonium chloride) (PDDA), was spin-coated on a tissue culture petridish and the micropatterns of the PDDA were formed by selective ion implantation through a pattern mask. The surface property of the implanted PDDA was investigated by using a surface profiler and FT-IR spectrometer. Cy3-labeled ssDNA was selectively immobilized on the PDDA patterns through ionic interaction and thus, well-defined ssDNA patterns were obtained.