• Title/Summary/Keyword: Natural DNA

Search Result 1,598, Processing Time 0.024 seconds

NMR Hydrogen Exchange Study of DNA Duplex Containing the Consensus Binding Site for Human MEIS1

  • Choi, Seo-Ree;Jin, Ho-seong;Seo, Yeo-Jin;Lee, Joon-Hwa
    • Journal of the Korean Magnetic Resonance Society
    • /
    • v.24 no.4
    • /
    • pp.117-122
    • /
    • 2020
  • Transcription factors are proteins that bind specific sites or elements in regulatory regions of DNA, known as promoters or enhancers, where they control the transcription or expression of target genes. MEIS1 protein is a DNA-binding domain present in human transcription factors and plays important roles in various biological functions. The hydrogen exchange rate constants of the imino protons were determined for the wild-type containing the consensus DNA-binding site for the MEIS1 and those of the mutant DNA duplexes using NMR spectroscopy. The G2A-, A3G- and C4T-mutant DNA duplexes lead to clear changes in thermal stabilities of these four consensus base pairs. These unique dynamic features of the four base pairs in the consensus 5'-TGAC-3' sequence might play crucial roles in the effective DNA binding of the MEIS1 protein.

Lipid Peroxidation Product-Mediated DNA Damage and Mutagenicity

  • Koh, Young-Ho;Yoon, Seon-Joo;Park, Jeen-Woo
    • BMB Reports
    • /
    • v.30 no.3
    • /
    • pp.188-193
    • /
    • 1997
  • Membrane lipid peroxidation processes yield products that may react with DNA to cause mutations. Lipid hydroperoxides from linoleic acid in the presence of transition metal ions caused strand breaks in plasmid DNA. DNA damage induced by reactive aldehydes known to be produced by decomposition of lipid hydroperoxides, such as 4-hydroxynonenal or rnalondialdehyde, was repaired by endonucleases and exonuclease III which resulted in the increase of single strand breaks in DNA. Lipid hydroperoxides as well as malondialdehyde and 4-hydroxynonenal also caused mutations in the pUC18 lacZ' gene when measured as a loss of ${\alpha}-cornplementation$. In conclusion. the lipid peroxidation could be an important intermediary event in DNA damage and mutation by oxidative stress.

  • PDF

Molecular Cloning of a cDNA Encoding a Ferritin Subunit from the Spider, Araneus ventricosus

  • Jin, Byung-Rea;Han, Ji-Hee;Kim, Seong-Ryul;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.4 no.2
    • /
    • pp.163-168
    • /
    • 2002
  • We report for the first time the cDNA sequence encoding a ferritin subunit from the spiders Araneus ventricosus. The complete cDNA sequence of A. ventricosus ferritin subunit comprised 516 bp with 172 amino acid residues. The A. ventricosus ferritin subunit cDNA contained a conserved iron responsive element sequence in the 5 untranslated region. An alignment of the deduced protein sequence of the A. ventricosus ferritin subunit gene to that of other heavy chain ferritin molecules showed that A. ventricosus ferritin subunit is most similar to the great pond snail, Lymnaea stagnalis, ferritin with 70.2% of protein sequence identity.

Unusual Mitochondrial DNA Polymorphism of the Blue Mussel (Mytilus edulis) Species Complex on the Southern Coast of Korea

  • Iksoo Kim;Byung-Yoon Min;Myung-Hee Yoon;Myong-Suk Yoo;Doh-Hoon Kim
    • Animal cells and systems
    • /
    • v.3 no.1
    • /
    • pp.79-87
    • /
    • 1999
  • Mitochondrial DNA (mtDNA) from 54 specimens of the blue mussel (Mytilus edulis) species complex sampled from the southern coast of Korea was assayed for polymorphism with a portion of the COIII gene (336 bp). Fifteen haplotypes were found. PAUP, one-step networks, and PHYLIP analyses revealed the presence of two clearly differentiated mitochondrial clades (termed clades B and E), separated by 3.6% of minimum sequence divergence. The distribution pattern of the species appears to be consistent with category II of the phylogeographic pattern sensu (Avise et al., 1987): the presence of two discontinuous and distinct mtDNA genotypes in the same geographic region. This unusual mitochondrial polymorphism was explained by the presence of the Mediterranean species, M. galloprovincialis, possessing mtDNA of both M. galloprovincialis and M. edulis.

  • PDF

Intraspecific variations of the Yam (Dioscorea alata L.) based on external morphology and DNA marker analysis

  • Chang, Kwang-Jin;Yoo, Ki-Oug;Park, Cheol-Ho;Lim, Hak-Tae;Michio Onjo;Park, Byoung-Jae
    • Plant Resources
    • /
    • v.3 no.3
    • /
    • pp.211-218
    • /
    • 2000
  • Intraspecific genetic relationship of 19 variation types of the Yam (Dioscorea alata) classified by their external morphological characteristics such as leaf and tuber shape were assessed by DNA using random and specific primer. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)$_4$ sequence]) had been used in PCR-amplification. Only 12 primers, however, were success in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0 %), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90. These DNA data were not matched well with those of morphological characters since they were divided into two major groups at the similarity coefficient value of 0.70. Therefore, Grouping of species into variation types by mainly morphological charactistics was suggested unreasonable.

  • PDF

A DNA Strand-Nicking Principle of a Higher Plant, Caesalpinia sappan

  • Mar, Woongchon;Lee, Hyun-Tai;Je, Kang-Hoon;Choi, Hye-Young;Seo, Eun-Kyoung
    • Archives of Pharmacal Research
    • /
    • v.26 no.2
    • /
    • pp.147-150
    • /
    • 2003
  • To find anticancer agents from higher plants, DNA strand-scission assay method was employed for bioassay-guided fractionation as well as for screening the crude extracts. During the screening, an ethyl acetate extracts of the heartwood of Caesalpinia sappan L. (Leguminosae) exhibited potent DNA strand-scission activity. Therefore, the ethyl acetate extracts of the dried heartwood of C. sappan was subjected to the bioassay-guided fractionation, which led to the isolation of a known compound, brazilin (1) as the active constituent. In addition, caesalpine J (2) was also isolated as an inactive constituent.

Rapid Isolation of Genomic DNA from Normal and Apoptotic Cells Using Magnetic Silica Resins

  • Park, Jee-Sun;Park, Jung-Hyun;Na, Shin-Young;Choe, Soo-Young;Choi, Sang-Nam;You, Kwan-Hee
    • Journal of Microbiology and Biotechnology
    • /
    • v.11 no.5
    • /
    • pp.890-894
    • /
    • 2001
  • The isolation of genomic DNA from mammalian cells is usually performed by cell lysis followed by protein digestion, extraction, and finally, ethanol precipitation of the chromosomal DNA. However, in the case of large sample numbers or when only small amounts of starting materials are available, such conventional methods are not efficient and are cumbersome to be applied. Some alternative methods have been described as well as having commercial DNA isolation kits to be available, nevertheless, there is room left for much improvement. In the present study, a novel method is introduced, where it simplifies conventional protocols by omitting some time-consuming steps such as protease incubation or DNA precipitation and its resuspension. Using paramagnetic silica resins, the genomic DNA was purified over a magnetic field, and the bound DNA was eluted with a low-salt buffer. The fidelity and effectiveness of this novel method was determined by using normal and apoptotic cells as a starting material and then compared to other protocols. The high speed and convenience along with its high efficiency in detecting apoptotic chromosomal DNA will prove this method to be an improved alternative in the isolation of genomic DNA from mammalian cells.

  • PDF

Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
    • /
    • v.3 no.1
    • /
    • pp.73-78
    • /
    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

  • PDF

Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

  • Park, Joung-Eon;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Cha, Jae-Ho;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
    • /
    • v.29 no.3
    • /
    • pp.121-131
    • /
    • 2001
  • This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

  • PDF

Free Radical Involvement in the DNA Damaging Activity of Fumonisin Bl

  • Lee, Wan-Hee;Lee, Kil-Soo
    • Toxicological Research
    • /
    • v.17 no.4
    • /
    • pp.249-253
    • /
    • 2001
  • Fumonisin B1, a mycotoxin, is thought to induce esophageal cancer in humans and apoptosis in animal cells by inhibiting ceramide synthase. Dumonisin Bl may also generate reactive oxygen species directly or indirectly, leading to DNA damage and lipid peroxidation. In this study, a DNA fragmentation assay, dichlorofluorescein (DCF) analysis, and single cell gel electrophoresis (SCGE) were used to investigate the involvement of cellular free radicals, specifically hydrogen peroxide, in the DNA damaging activity of fumonisin B1. From an in vitro DNA fragmentation assay, E. coli DNA, damage by fumonisin Bl was increased by the addition of superxide dismutase (SOD) and decreased by catalase. SCGE and DCF analysis in vivo showed that the nuclear DNA damage and intracellular free radicals in cultured rat hepatocytes treated with fumonisin B1 were increased with the concentration of fumonisin Bl . DNA damage and free radical generation were inhibited by the addition of catalase. Fumonisin Bl , in the presence of SOD, produces hydrogen peroxide causing oxidative DNA damage and protein malfunction, leading to genotoxicity and cytotoxicity of the toxin.

  • PDF