• Title/Summary/Keyword: Natural DNA

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Identification of a Tandemly Repented DNA Sequence Using Combined RAPD and FISH in Welsh Onion (Allium fistulosum)

  • Bong Bo Seo;Geum Sook Do;Seon Hee Lee
    • Animal cells and systems
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    • v.3 no.1
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    • pp.69-72
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    • 1999
  • A tandemly repeated DNA sequence was identified and characterized y the combined RAPD and FISH data from a total genomic DNA of Welsh onion (Allium fistulosum). A clone containing this repeating sequence was selected and sequenced. This repeating unit of 314 bp inserted into pAf 072 contained 54.1% adenine and thymine residues, and showed the primer sequence used, 5'-GAAACGGGTG-3', in both terminals of the sequence. Fluorescence in situ hybridization using this tandemly repeated sequence as a probe indicated that the detected sites were coincident with the major C-banded constitutive heterochromatin in the terminal regions of both arms of all 6 chromosomes.

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Mitochondrial DNA Analysis of the Small Yellow Croaker (Pseudosciaena polyactis Bleeker) in the Yellow Sea (황해산 참조기 (Pseudosciaena polyactis Bleeker)의 mitochondrial DNA 분석)

  • HWANG Gyu-Lin;LEE Yong-Chul;CHANG Chung-Soon;HUE Hoi-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.613-619
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    • 1994
  • To investigate the population differences of small yellow croaker (Pseudosciaena polyactis BLEEKER) in the Yellow Sea, five catching sites (three from China; Zoushan, Shanghai and Qingdao, two from Korea; Inchon and Mokpo) were selected for sampling. The populations of small yellow croaker from all five catching sites were investigated to analyze their mtDNA's restriction fragment length polymorphism (RFLP) using 18 kinds of restriction enzymes. The average molecular size of the entire mtDNA was estimated at $16.9{\pm}0.6\;kb$. According to the results of RFLP analysis, a total of 40 restriction sites were identified in every population surveyed and the overall cleavage patterns of mtDNA, based on the RFLP, showed similar tendencies. However, the five restriction enzymes such as ApaI, EcoRI, PstI, SmaI and SstII showed slightly different cleavage patterns which could have resulted from individual variations between the populations of Korea and China.

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Glycation of Copper, Zinc-Superoxide Dismutase and its Effect on the Thiol-Metal Catalyzed Oxidation Mediated DNA Damage

  • Park, Jeen-Woo;Lee, Soo-Min
    • BMB Reports
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    • v.28 no.3
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    • pp.249-253
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    • 1995
  • The nonenzymatic glycation of copper, zinc-superoxide dismutase (Cu,Zn-SOD) led to inactivation and fragmentation of the enzyme. The glycated Cu,zn-SOD was isolated by boronate affinity chromatography. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA and the generation of strand breaks in pBhiescript plasmid DNA by a metal-catalyzed oxidation (MCO) system composed of $Fe^{3+}$, $O_2$, and glutathione (GSH) as an electron donor was enhanced more effectively by the glycated CU,Zn-SOD than by the nonglycated enzyme. The capacity of glycated Cu,Zn-SOD to enhance damage to DNA was inhibited by diethylenetriaminepentaacetic acid (DETAPAC), azide, mannitol, and catalase. These results indicated that incubation of glycated CU,Zn-SOD with GSH-MCO may result in a release of $Cu^{2+}$ from the enzyme. The released $Cu^{2+}$ then likely participated in a Fenton-type reaction to produce hydroxyl radicals, which may cause the enhancement of DNA damage.

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Extension of a 5'- or 3'-end Genomic DNA Sequence by a Single PCR Amplification

  • Jeon, Taeck J.
    • Journal of Integrative Natural Science
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    • v.1 no.3
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    • pp.230-233
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    • 2008
  • A simple and rapid method is described for extending the 5'- or 3'-end genomic sequence of a known partial sequence by only a single round of PCR. This method involves digesting and ligating genomic and plasmid DNAs, and amplifying the 5'-upstream or 3'-end downstream sequence of the known DNA sequence, using two primers, one gene specific and the other plasmid specific. A single round of PCR amplification is sufficient to produce gene-specific bands detectable in gels. By using this approach, 5'-end genomic sequence of the D-amoeba sams gene was extended.

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Chiral Separation with DNA-Polyion Complex Membranes

  • Yoshikawa, Masakazu;Maruhashi, Motokazu;Ogata, Naoya
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.353-353
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    • 2006
  • Deoxyribonucelic acid (DNA) molecules have a huge molecular weight so that DNA was reported to be a promising natural polymer to give durable films. Among many applications of DNA, the authors focused their attention on separation membranes derived from DNA because membranes will play an important role in environmental and energy related processes. DNA-polyion complex membranes were prepared from DNA and corresponding polycations. The DNA-polyion complex membranes showed chiral separation ability toward racemic amino acid mixtures.

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Molecular Identification and Chemical Analysis of Aconiti Kusnezoffii Tuber on the Domestic Markets (국내 시장에서 유통되는 초오의 DNA 감별과 화학적 분석)

  • Jang, Hyeri;Joe, Kyeong-Hwa;Song, Kwangho;Lee, Kyoung Jin;Park, Sait Byul;Lee, Chaemin;Ha, In Jin;Lee, Kyungjin;Suh, Youngbae;Kim, Yeong Shik
    • Korean Journal of Pharmacognosy
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    • v.49 no.2
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    • pp.145-154
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    • 2018
  • Aconiti Kusnezoffii Tuber has been traditionally used to treat the symptoms of rheumatoid arthritis and joint pain. The main constituents are diterpenoid alkaloids such as benzoylmesaconine, benzoylaconine, mesaconitine, aconitine, and hypaconitine. In Korea, Aconiti Kusnezoffii Tuber is officially defined as the tubers of Aconitum kusnezoffii Reichb., A. ciliare Decasisne, and A. triphyllum Nakai. On the other hand, only the tuber of A. kusnezoffii is to be used in China. In order to identify the botanical origin of Aconiti Kusnezoffii Tuber circulated in Korea, we analyzed 24 samples of Aconiti Kusnezoffii Tuber obtained from local markets for comparative DNA analysis. The sequence analysis of nrRNA ITS 1 was useful to distinguish Aconitum species and revealed that the roots of A. karakolicum were circulated in Korean markets without discretion. HPLC quantitative analysis showed that aconitine was detected at the highest amount in A. karakolicum. Authentic diterpenoid alkaloids were coinjected for quantification of aconitine-type ingredients. All data were statistically grouped by Principal Component Analysis (PCA). This study suggests that both molecular and chemical analyses should be utilized for the standardization and the quality control for Aconiti Kusnezoffii Tuber.

The Genetic Diversity of Bacterial Communities in the Groundwater (지하수 세균 군집의 유전적 다양성)

  • 김여원;민병례;최영길
    • Korean Journal of Environmental Biology
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    • v.18 no.1
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    • pp.53-61
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    • 2000
  • In order to characterize the genetic diversity of bacterial community in groundwater, samples were collected from used for drinking water and polluted with heavy metal wastewater in Seoul city and natural cave of Kangwondo. The DNA was amplified with 165 rDNA-based primers by use of the PCR, and then analysed ARDRA (amplified ribosomal DNA restriction analysis). Restriction endonuclease analysis patterns of amplified 165 rDNA in drinking water and wastewater relatively showed high genetic diversity in situ and drinking groundwater. The number of DNA fragments varied with in situ and drinking water. This method of ARDRA of bacterial communities in groundwater could be used for a quick assessment of genotypic changes between different locations reflecting different environmental conditions and the diversity reflected pollution of groundwater (natural cave water>drinking water>waste water, as in order of grade). [Genetic diversity, Groundwater, 165 rDNA, PCR, ARDRA].

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CpG-DNA induces bacteria-reactive IgM enhancing phagocytic activity against Staphylococcus aureus infection

  • Kim, Te Ha;Kim, Dongbum;Lee, Heesu;Kwak, Min Hyung;Park, Sangkyu;Lee, Younghee;Kwon, Hyung-Joo
    • BMB Reports
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    • v.52 no.11
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    • pp.635-640
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    • 2019
  • CpG-DNA triggers the proliferation and differentiation of B cells which results in the increased production of antibodies. The presence of bacteria-reactive IgM in normal serum was reported; however, the relevance of CpG-DNA with the production of bacteria-reactive IgM has not been investigated. Here, we proved the function of CpG-DNA for the production of bacteria-reactive IgM. CpG-DNA administration led to increased production of bacteria-reactive IgM both in the peritoneal fluid and serum through TLR9 signaling pathway. When we stimulated B cells with CpG-DNA, production of bacteria-reactive IgM was reproduced in vitro. We established a bacteria-reactive monoclonal IgM antibody using CpG-DNA stimulated-peritoneal B cells. The monoclonal IgM antibody enhanced the phagocytic activity of RAW 264.7 cells against S. aureus MW2 infection. Therefore, we suggest that CpG-DNA enhances the antibacterial activity of the immune system by triggering the production of bacteria-reactive IgM. We also suggest the possible application of the antibodies for the treatment of antibiotics-resistant bacterial infections.

DNA Fingerprinting Analysis of Natural Monument Gingko Trees Using Microsatellite Markers (Microsatellite 마커를 이용한 은행나무 천연기념물의 DNA 지문 분석)

  • Lee, Jei-Wan;Lee, Min-Woo;Ahn, Ji-Young;Hong, Kyung-Nak;Baek, Seung-Hoon;Kim, Sang-Chul
    • Journal of Korean Society of Forest Science
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    • v.106 no.4
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    • pp.408-416
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    • 2017
  • This study describes DNA fingerprinting analysis of twenty-three natural monument individuals of Ginkgo biloba using eight microsatellite markers. The average number of observed alleles was 6.875, and the expected heterozygosity and the observed heterozygosity were 0.711 and 0.710, respectively. This results were similar to those of the previous studies on Ginkgo trees analyzed by same markers in China and Japan. PIC value and PD were calculated at 0.677 and 0.9999 respectively, indicating a high individual identification efficiency. In fact, all of the natural monument ginkgo trees and additionally analyzed thirteen general ginkgo tress were identified by genotype comparison. PI and PD calculated in three markers (Ging06, Gb60, Gb61) with the highest PIC values calculated in natural monument ginkgo trees were $8.045{\times}10^{-5}$ and 99.99%, respectively. Thus, these three markers could be preferentially used in DNA fingerprinting for identifying ginkgo tree individuals. The results in this study will be useful for management of natural monument ginkgo trees, proliferation of their progeny and genetic identification of individuals selected in breeding process.

Molecular Cloning of a cDNA Encoding Putative Calreticulin from the Silkworm, Bombyx mori

  • Kim, Seong-Ryul;Lee, Kwang-Sik;Kim, Iksoo;Kang, Seok-Woo;Nho, Si-Kab;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.1
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    • pp.93-97
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    • 2003
  • We describe here the cloning of a cDNA encoding putative calreticulin (CRT) from the silkworm, Bombyx mori. The CRT cDNA comprised of 1,194 bp encoding 398 amino acid residues. B. mori. CRT has a HDEL sequence at the end of the C-domain. The B. morl, CRT showed 88% protein sequence identity to the G. mellonella CRT, 71 % to A. aegypti CRT, and 63% to H. sapiens CRT, Phylogenetic analysis revealed that the deduced amino acid sequences of the B. mori CRT formed a highly inclusive subgroup with other insect CRTs. Northern blot analysis exhibited an expression of the B. mori CRT gene in the fat body, evidencing the fat body as a major site for CRT synthesis.