• Title/Summary/Keyword: DNA.

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Mitochondrial DNA Analysis in Fusants of Ganoderma lucidum and Lentinus edodes (영지버섯과 표고버섯 원형질 융합체의 미토콘드리아 DNA 검색)

  • 최은주;정영자;이영재;김병각;현진원
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.199-204
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    • 2002
  • It has been known that Ganoderma lucidum and Lentinus edodes have anticancer activity and immune enhancing activity. These two mushrooms were grown in liquid culture and harvested. From these mycelia, DNA was isolated and EtBr-CsCl density gradient ultracentrifugation was performed to purify it further. Then mitochondrial DNA was isolated by bisbenzimide-CsCl density ultracentrifugaton. Mitochondrial DNA of Ganoderma lucidum was digested by restriction enzymes, EcoR I, Hind Ⅲ, and Pst I, then electrophoresed. It showed 12, 22, 4 fragments. Mitochondrial DNA of Lentinus edodes was digested by EcoR I. Electric pattern showed 6 fragments. 4 fragments had appeared by Pst 1 digested mitochondrial DNA. Hind ill couldn't digest mitochondrial DNA of Lentinus edodes. Mitochondrial DNA of fusants was isolated to compare to those of parents. The results showed that fusant P₂S₄has new, recombined mitochondrial DNA. But P₂S₄had the same DNA that Ganoderma lucidum had.

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DNA Chip using Single Stranded Large Circular DNA: Low Background and Stronger Signal Intensity

  • Park, Jong-Gu
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.75-84
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    • 2004
  • Massive identification of differentially expressed patterns has been used as a tool to detect genes that are involved in disease related process. We employed circular single stranded sense molecules as probe DNA for a DNA chip. The circular single stranded DNAs derived from 1,152 unigene cDNA clones were purified in a high throughput mode from the culture supernatant of bacterial transformants containing recombinant phagemids and arrayed onto silanized slide glasses. The DNA chip was examined for its utility in detection of differential expression profile by using cDNA hybridization. Hybridization of the single stranded probe DNA were performed with Cy3- or Cy5-labeled target cDNA preparations at $60^\circ$C. Dot scanning performed with the hybridized slide showed 29 up-regulated and 6 down-regulated genes in a cancerous liver tissue when compared to those of adjacent noncancerous liver tissue. These results indicate that the circular single stranded sense molecules can be employed as probe DNA of arrays in order to obtain a precious panel of differentially expressed genes.

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Effective Family Shuffling Method Using Complementary DNA Fragments Produced by S1 Nuclease

  • Hong, Soon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.2004-2007
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    • 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%.

Gamma-ray Induced DNA Repair Synthesis in Relation to Chromosome Exchanges in Mammalian Cells in Vitro (哺乳動物細胞에 있어 감마線에 의한 DNA 回復合成과 染色體交換과의 聯關性)

  • Park, Sang-Dai
    • The Korean Journal of Zoology
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    • v.18 no.1
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    • pp.41-49
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    • 1975
  • Dose response and time dependence of DNA repair synthesis were investigated to determine the possible relationship between DNA repair synthesis and chromosome exchanges in $\\gamma$-ray irradiated BHK-21 and KB cell lines. DNA repair synthesis induced by $\\gamma$-ray was dose dependent up to 5kR, then leveling off occurred until 50 kR was reached. Time dependence of DNA repair synthesis was continued for up to 1$\\sim$2 hours after irradiation although the initial dose responses were cell line specific. Chromosome exchanges induced by $\\gamma$-ray showed different radiosensitivities in these cell lines and did not show a correlation with the DNA repair synthesis.

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HBV Polymerase Residues $Asp^{429}$ and $Asp^{551}$, Invariant at Motifs A and C are Essential to DNA Binding

  • Kim, Youn-Hee;Hong, Young-Bin;Jung, Gu-Hung
    • BMB Reports
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    • v.31 no.5
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    • pp.498-502
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    • 1998
  • HBV polymerase shares several regions of amino acid homology with other DNA-directed and RNA-directed polymerases. The amino acid residues $Asp^{429}$, $Gly^{518}$, $Asp^{551}$, $Lys^{585}$, and $Gly^{641}$ in the conserved motifs A, B', C, D, and E in the polymerase domain of HBV polymerase were mutated to alanine or histidine by in vitro site-directed mutagenesis. Those mutants were overexpressed, purified, and analyzed against DNA-dependent DNA polymerase activity and affinity for DNA binding. All those mutants did not show DNA-dependent DNA polymerase activities indicating that those five amino acid residues are all critical in DNA polymerase activity. South-Western analysis shows that amino acid residues $ASp^{429}$ and $ASp^{551}$ are essential to DNA binding, and $Gly^{318}$ and $Gly^{585}$ also affect DNA binding to a certain extent.

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Development of New DNA Chip and Genome Detection Using an Indicator-free Target DNA (비수식화 DNA를 이용한 유전자 검출 및 새로운 DNA칩의 개발)

  • Park, Yong-Sung;Park, Dae-Hee;Kwon, Young-Soo;Tomoji Kawai
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.52 no.8
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    • pp.365-370
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    • 2003
  • This research aims to develop an indicator-free DNA chip using micro-fabrication technology. At first, we fabricated a DNA microarray by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the gold electrodes. Then indicator-free target DNA was hybridized by an electrical force and measured electrochemically in potassium ferricyanide solution. Redox peak of cyclic-voltammogram showed a difference between target DNA and mismatched DNA in an anodic peak current. Therefore, it is able to detect various genes electrochemically after immobilization of various probe DNAs and hybridization of indicator-free DNA on the electrodes simultaneously It suggested that this DNA chip could recognize the sequence specific genes.

Effects of Ginseng Saponin on DNA Strand Breaks and Replication Inhibition by Benzo(a)Pyrene in CHO-Kl Cells (Benzo(a)Pyrene 유발 DNA 상해 및 복제 억제에 미치는 인삼사포닌의 영향)

  • Park, Jin-Kyu;Park, Ki-Hyun
    • Journal of Ginseng Research
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    • v.16 no.3
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    • pp.210-216
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    • 1992
  • The effect of saponin extracted from Panax grneng CA Meyer on DNA repair and replicative DNA synthesis were examined in CHO-Kl cells cotreated with benzo(a)pyrene and rat liver S-15 fraction. The DNA strand breaks inititated by benzo(a)pyrene metabolites were measured by alkaline election technique. The addition of ginseng saponin to the culture media resulted in decrease of benzo(a)pyrene-induced DNA strand breaks, and restored the suppressed-semiconservative-DNA-synthesis by the carcinogen. DNA repair synthesis in the damaged cells was also elevated by the ginseng treatment when the repairing activites were measured for the (3H)-thymidine incorporation into the carcinogen damaged cellular DNk Comparative analysis of DNA-adduces of benzo(a)pyrene metabortes in microsomes suggested that ginseng saponin treatment in rats reduced the formation of electrophilic metabolites of benzo (a)-pyrene in the rat liver microsomes.

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Effect of saltss on the entrapment of calf thymus DNA into liposomes

  • Kim, Chong-Kook;Lee, Beom-Jin
    • Archives of Pharmacal Research
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    • v.10 no.2
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    • pp.110-114
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    • 1987
  • To correlate the conformational changes of DNA (Calf Thymus) with entrapment of DNA into liposomes, the effect of ions ($Na^+$, $Mg^{++}$on the entrapment of calf thymus DNA into liposomes was investigated. The effect of divalent ion ($Mg^{++}$ on the structural changes of DNA indicated by decrease of observed ellipticity at 274 nm and nonspecific binding of DNA to lipid bilayers was greater than monovalent ion ($\Na^+$). But the efficiency of DNA encapsulated was not altered. These results show that entrapment of DNA into liposomes is not due to nonspecific binding and structural changes because of electrostatic forces but to mechanical capture of DNA by the internal aqueous space of liposomes although divalent ion contributes large structural changes and more nonspecific association of DNA with liposomes due to strong charges.

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Implementation of GA Processor for Efficient Sequence Generation (효율적인 DNA 서열 생성을 위한 진화연산 프로세서 구현)

  • Jeon, Sung-Mo;Kim, Tae-Seon;Lee, Chong-Ho
    • Proceedings of the KIEE Conference
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    • 2003.11c
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    • pp.376-379
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    • 2003
  • DNA computing based DNA sequence Is operated through the biology experiment. Biology experiment used as operator causes illegal reactions through shifted hybridization, mismatched hybridization, undesired hybridization of the DNA sequence. So, it is essential to design DNA sequence to minimize the potential errors. This paper proposes method of the DNA sequence generation based evolutionary operation processor. Genetic algorithm was used for evolutionary operation and extra hardware, namely genetic algorithm processor was implemented for solving repeated evolutionary process that causes much computation time. To show efficiency of the Proposed processor, excellent result is confirmed by comparing between fitness of the DNA sequence formed randomly and DNA sequence formed by genetic algorithm processor. Proposed genetic algorithm processor can reduce the time and expense for preparing DNA sequence that is essential in DNA computing. Also it can apply design of the oligomer for development of the DNA chip or oligo chip.

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DNA and DNA-CTMA composite thin films embedded with carboxyl group-modified multi-walled carbon nanotubes

  • Dugasani, Sreekantha Reddy;Gnapareddy, Bramaramba;Kesama, Mallikarjuna Reddy;Ha, Tai Hwan;Park, Sung Ha
    • Journal of Industrial and Engineering Chemistry
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    • v.68
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    • pp.79-86
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    • 2018
  • Although the intrinsic characteristics of DNA molecules and carbon nanotubes (CNT) are well known, fabrication methods and physical characteristics of CNT-embedded DNA thin films are rarely investigated. We report the construction and characterization of carboxyl (-COOH) group-modified multi-walled carbon nanotube (MWCNT-COOH)-embedded DNA and cetyltrimethyl-ammonium chloride-modified DNA (DNA-CTMA) composite thin films. Here, we examine the structural, compositional, chemical, spectroscopic, and electrical characteristics of DNA and DNA-CTMA thin films consisting of various concentrations of MWCNT-COOH. The MWCNT-COOH-embedded DNA and DNA-CTMA composite thin films may offer a platform for developing novel optoelectronics, energy harvesting, and sensing applications in physical, chemical, and biological sciences.