• Title/Summary/Keyword: DNA length

Search Result 1,316, Processing Time 0.029 seconds

Mechanism study on DNA damage and Apoptosis induced by heak shock using Comet Assay

  • Seo, Young-Rok;Han, Sung-Sik;Kim, L. O′Neill;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
    • /
    • 1997.12a
    • /
    • pp.101-101
    • /
    • 1997
  • Comet assay, single cell gel electrophoresis has been known as useful, rapid, simple, visual, and sensitive technique for measuring the DNA breakage in mammalian ce1ls. For evaluation of DNA damage using comet assay, early studies reported a change in comet length and intensity with DNA damage using simple visual technique, such as fluorescence microscopy with eyespiece. In recent, some workers are observing and analyzing nucleotide of comets using quantitative fluorescence image analysis system to estimate 'tail moment', which is defined as the product of the tail length and the fraction of total DNA in tail. Our laboratory also adopted the image analysis software for qualification. In addition, many of the practical features of comet assay render it potentially attractive as useful tool for molecular toxicology and carcinogenesis, because the system is already showing considerable promise as rapid predictor in both in vitro and in vivo experimental designs. Recently, the comet assay becomes a attractive technique to study of apoptosis, because apoptotic fragmentation of nuclear DNA into nucleosomal sizes can be evaluated by the comet assay. So, we attempted to apply the comet assay to studying the effect of various stress on the apoptosis-sensitive cell lines. Particularly, focusing on the hyperthermic apoptosis, we could find that heat shock(44˚C for 60 minutes) was sufficient to induced apoptosis in these cell lines. But using the highly sensitive comet assay, we could not detect DNA breaks immediately after heat shock.

  • PDF

Karyotype Analysis and Physical Mapping of rDNAs in Bupleurum longeradiatum (개시호 (Bupleurum longeradiatum)의 핵형분석과 rDNAs의 Physical Mapping)

  • Koo, Dal-Hoe;Seong, Nak-Sul;Seong, Jong-Suk;Bang, Kyong-Hwan;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
    • /
    • v.11 no.5
    • /
    • pp.402-407
    • /
    • 2003
  • Karyotype analysis and chromosomal localization of 5S and 45S rDNAs using multi-color fluorescence in situ hybridization (McFISH) technique were carried out in Bupleurum longeradiatum. Somatic metaphase chromosome number was 2n=12. Karyotype was composed of three pairs of metacentrics (No.3, 4 and 6) and three pairs of submetacentrics (No. 1, 2 and 5). The length of somatic prometaphase chromosomes ranges from 2.55 to $5.05{\mu}m$ with total length of $18.15\;{\mu}m$. In FISH experiment, one pair of 5S rDNA signals was detected on the pericentromeric region of chromosome 4 and one pair of 45S rDNA signals was detected on the telomeric region of chromosome 2.

Identification Characteristics of Irradiated Dried Red Pepper during Storage by Analysis of Thermoluminescence, DNA Comet, and DEFT/APC (Thermoluminescence, DNA Comet 및 DEFT/APC 분석에 의한 방사선처리 건고추의 저장 중 검지 특성)

  • Kim, Byeong-Keun;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
    • /
    • v.36 no.6
    • /
    • pp.851-856
    • /
    • 2004
  • Minerals separated from irradiated dried red pepper (whole) at 2.5 kGy or higher showed typical thermoluminescence (TL) glow curves ($TL_1$) at around $150^{\circ}C$, which increased with irradiation dose. The TL ratio ($TL_1/TL_2$) through re-irradiation step at 1 kGy enhanced reliability of TL identification results. DNA comet assay indicated that the intact cell was observed in non-irradiated pepper (seed), while some long tails were found in irradiated ones, showing relationship between irradiation dose and tail length. Log DEPT/APC values increased in proportion to irradiation doses in powdered and whole peppers. Based on overall results, irradiated dried red peppers could be screened using DNA comet assay or log DEFT/APC, and moreover the identification results were verified by TL analysis.

Systematic Studies on Korean Rodents: VI. Analysis of Morphometric Characters, Chromosomal Karyotypes and Mitochondrial DNA in Two Species of Genus Rattus (한국에 서식하고 있는 설치류의 계통분류학적 연구: 6. 집쥐속 2종의 형태학적 형질, 염색체 핵형 및 미토콘디리아 DNA의 분석)

  • 고홍선
    • Animal Systematics, Evolution and Diversity
    • /
    • v.8 no.2
    • /
    • pp.231-242
    • /
    • 1992
  • Samples of two species of genus Ratts(black rat, Rattus rasttus Linnaeus; common rat, Rattus norvegicus Berkenhaut) in Korea were trapped and their 31 morphometric charcters were analyzed statistically in order to determine the range of geographic variation within each species and the interspecific differences. In addition, chromosomal G-bands and C-bands were compared and the fragment patterns of mtDNA resulted from the digestion with restriction enzymes were also analyzed. Samples of black rats from six localities in Korea were similar with one another in their morphometric characters: in head and body length, length of tail vertebrae, conventional karyotype and C-bands, they are comparable to Rattus rattus tanezumi in Japan. Specimens of common rats from seven localities in Korea were similar with one another in their morphometric characters: in conventional karyotype, they are comparable to Rattus norvegicus caraco in eastern Asia. Common rats differ from black rats in their morphometric characteris, chromosomal karyotypes and mtDNA. It is confirmed that correct species name of black rat in Korea is Rattus rattus tanezumi Tempminck: species name of common rat in Korea is Rattus norvegicus caraco Pallas: the common rat is a species, which is distinct from the black rat.

  • PDF

Effects of Gamma-ray Irradiation on Growth Characteristics and DNA Damage in Licorice (Glycyrrhiza uralensis) (감마선 조사가 감초(Glycyrrhiza uralensis)의 초기 생육 및 DNA 손상에 미치는 영향)

  • Ryu, Jaihyunk;Im, Seung Bin;Kim, Dong Sub;Ahn, Joon-Woo;Kim, Jin-Baek;Kim, Sang Hoon;Kang, Si-Yong
    • Journal of Radiation Industry
    • /
    • v.8 no.2
    • /
    • pp.89-95
    • /
    • 2014
  • This study was conducted to determine the optimal dose of gamma-ray on the growth and nucleus DNA damage for mutation breeding in licorice. Gamma-rays irradiated to dry seeds with various doses (0 to 1000 Gy). Significant decreases in germination rate (%), survival rate (%) and growth characteristics (plant height, number of leaves, root length and fresh weight) were observed by dose of increased. $LD_{50}$ (lethal dose) was approximately 400 Gy to 500 Gy. Also, reduction doses ($RD_{50}$) of plant height, number of leaves, root length and flash weight were 428 Gy, 760 Gy, 363 Gy and 334 Gy, respectively. It is supplest that the optimal dose of gamma irradiation for licorice mutation induction might be about 400 Gy in this study. We also conducted comet assay to observe nucleus DNA damage due to gamma irradiation. In comet assay, a clear difference was identified over 300 Gy treatments. With increasing doses of gamma-ray in the range of 100 to 1000 Gy, the rate of head DNA was decreased significantly from 92.88% to 73.09%. Tail length(${\mu}m$) was increased as the dose of increased over 300 Gy. Growth characteristics (Germination rate, Survival rate, plant height, number of leaves, root length and fresh weight) were highly negatively ($P{\leq}0.01$) correlated with dose. While the tail length was highly positively ($P{\leq}0.01$) correlated with dose.

Genetic Analysis study of Sasang Constitution Classification by RFLP (RFLP법을 이용한 사상체질의 유전적 분석 연구)

  • Cho, Dong-Wuk;Cho, Hwang-Sung
    • Journal of The Association for Neo Medicine
    • /
    • v.2 no.1
    • /
    • pp.25-33
    • /
    • 1997
  • In Sasang medicine, humen are classified into four constitutions which are Taeyang, Soyang. Taeum and Soum. Depending on each different constitution, the clinical and pharmacological application for the same disease might be different. In this study, genomic DNA of different constitutions(Taeum, Soyang and Soum) were analyzed by Restriction Fragment Length Polymorphism(RFLP) to provide scientific and objective references for Sasang classification. The DNA polymorphism for each constitution detected as differences in the length of DNA fragments, after digestion with restriction enzyme Hae III, was investigated using YNH24 as DNA probe. The allele size of Taeum, Soyang and Soum group detected by YNH24 ranged from 1.3 to 3.8 kb, 1.5 to 3.9 kb and 1.3 to 4.6 kb, respectively. However, the allele size distribution of YNH24 loci of different constitutions was shown to be too variable to be classified as 3 different constitution groups investigated. For further study, it is suggested that the number of each constitution samples for RFLP analysis should be increased and statistical analysis of the allele size distribution should be carried out.

  • PDF

Comparison of Terminal-restriction Fragment Length Polymorphism (T-RFLP) Analysis and Sequencing of 16S rDNA Clones in marine sediments

  • Lee Jung-Hyun
    • Proceedings of the Microbiological Society of Korea Conference
    • /
    • 2002.10a
    • /
    • pp.15-21
    • /
    • 2002
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis has been optimized by using in vitro model community composed of genomic DNAs of known bacterial strains and has been applied to assess the bacterial community structure in marine sediments. The specific fluorescence-labeled terminal restriction fragments (T-RFs) between 39 and 839 base long specifying each strain were precisely measured for known bacterial strains. The addition of a co-solvent (dimethylsulfoxide or glycerol) into PCR reactions has reduced differential PCR amplification. Comparative bacterial community structure was investigated for pristine and polluted sediments. A complex T-RFLP pattern showing complex bacterial community structure was obtained in the pristine sediment, whereas simple T-RFLP pattern (low bacterial diversity) was shown in polluted sediments where caged aquaculture has been conducted for several years. The results of T-RFLP analysis were compared with that of cloning and sequencing 16S rDNA clones from the same sediments. Sequence analysis of 16S rDNA clones (72) of the pristine sediment revealed a diverse collection of lineages, largely of the class Proteobacteria ($6\%$ alpha subdivision, $46\%$ gamma subdivision, $13\%$ delta subdivision, and $3\%$ epsilon subdivision), Nitrospina $(8\%)$, high G+C gram positive $(8\%)$, Verrucomicrobia $(7\%)$, and Planctomycetes $(6\%)$. In the contaminated sediments, 17 $(59\%)$ of the 16S rDNA clones (29) were related to Campylobacter and symbiont of Rimicaris exoculata belonging to epsilon subdivision of Proteobacteria. The results obtained indicated that T-RFLP analysis is a rapid and precise technique for comparative bacterial community analysis.

  • PDF

Zero-Length Crosslinking Study on Interactions of TBP, GAL4-AH, and TFIIB in the Preinitiation Complex (전사개시전 복합체에서 TBP, GAL4-AH, TFIIB의 상호작용에 대한 Zero-Length Crosslinking 실험)

  • 권혁만
    • The Korean Journal of Zoology
    • /
    • v.39 no.4
    • /
    • pp.393-399
    • /
    • 1996
  • A zero-Iength croessinking procedure for studying protein-protein interactlons in preinitiation complex has heen developed. Preinltiadon complexes were assembled with immobilized DNA templates coupled to metal beads. Pudfied complexes were dIrectly crosslinked by 1-ethyl-3.(3-dimethylaminopropyl)carbodlimide (EDC). The reaction was stopped by addition of $\beta$-mercaptoethanol, and the complexes were isolated from EDC immedIately. An appllcatlion of this method with a preinltlation complex assembled with TBP, TFIIB, and GAL4-AH demonstrated that ThP dIrectly interacts with GAL4-AH and TFIIB in the prelnitlatlon complex. However, croeslinked produd between GAL4-AH and ThilB was not observed. These resutts lndlcate that GAL4-AH does not stably Interact with TFIIB In the GAL4-AH-TFIIB-TBP-DNA prelnitlatlon complex.

  • PDF

RFLP Analysis of the mtDNA COI Region in Four Abalone Species

  • Park, Choul-Ji;Kijima, Akihiro
    • Fisheries and Aquatic Sciences
    • /
    • v.9 no.3
    • /
    • pp.101-106
    • /
    • 2006
  • The cytochrome c oxidase subunit I (COI) gene region of mitochondrial DNA (mtDNA) was examined in four abalone species to estimate its utility as a genetic marker using restriction fragment length polymorphism (RFLP) analysis. The utility was evaluated in terms of genetic divergence and relationships among Haliotis discus hannai, H. rufescens, H. rubra, and H. midae in both hemispheres of the world. There was clear genetic divergence in the mtDNA COI region between all pairs of the four species. Moreover, relationships among the abalone species were reflected in their geographical distributions and morphological characteristics. Therefore, RFLP analysis of the mtDNA COI region is a suitable genetic marker for the estimation of genetic divergence and relationships among abalone species. However, it is not effective for the evaluation of genetic differences within abalone species.

DNA Length Dependent Photocurrent of Diketopyrrolopyrrole Aggregates Constructed with DNA

  • Nakamura, Mitsunobu;Tsuto, Koji;Takada, Tadao;Yamana, Kazushige
    • Rapid Communication in Photoscience
    • /
    • v.3 no.4
    • /
    • pp.67-69
    • /
    • 2014
  • Bis(2-thienyl)-diketopyrrolopyrrole having two $Zn^{II}$-cylcens (DPPCy) was synthesized. DPP-aggregates were constructed by self-organization of DPPCy and $dT_n$-DNAs. In the presence of L-ascorbic acid as an electron sacrifice reagent, the DPP aggregates immobilized on a gold electrode exhibit good anodic photocurrent responses as well as cathodic photocurrent responses in the presence of methyl viologen. The anodic photocurrent responses depend on the DNA lengths because of the formation of uniform DPP-aggregates corresponding to the DNA lengths. The present results show that photocurrent responses of the DPP-aggregates can be controlled by DNA lengths and electron sacrifice reagents.