• Title/Summary/Keyword: Dna2

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Application of DNA Probe Method for Detection of 2,4-Dichlorophenoxyacetic Acid Degrading Bacteria in Soil (DNA Probes에 의한 토양의 이사디 (2,4-D) 분해세균의 검출)

  • Ka, Jong-Ok
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.403-408
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    • 1996
  • Total bacterial community DNA, which was extracted from microcosm soil and field soil after 2,4-D amendments, was analyzed on Southern blots, using the tfdA gene probe derived from plasmid pJP4 and the Spa probe from Sphingomonas paucimobilis. Southern blot analyses with total bacterial DNA extracted from soils Inoculated with Pseudomonas cepacia/pJP4 revealed that DNA probe method could detect the 2,4-D degrading bacteria down to $10^5\;cells/g$ dry soil. In the microcosm experiment, there was a good correlation between 2,4-D degradation and banding patterns in hybridization analyses performed after each 2,4-D treatment using the two probes. When bacterial DNA extracted from microcosm soil was hybridized with the Spa probe, a change in the position of hybrid bands was observed over time in a Southern blot, suggesting that population change or possibly genetic rearrangement in 2,4-D degrading microbial populations occurred in this soil. With the Spa probe, one hybrid DNA band was persistently observed throughout the five 2,4-D additions. When bacterial DNA isolated from the field soil was probed with the tfdA and Spa, strong hybridization signal was observed in the 100 ppm-treated subplot, weak signal In the 10 ppm-treated subplot, and no significant signal in the 1 ppm-treated and control subplots. The data show that DNA probe analyses were capable of detecting and discriminating the indigenous 2,4-D degrading microbial populations in soil amended with 2,4-D under laboratory and field conditions.

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Synthesis and Characterization of DNA-mediated Gold Nanoparticles by Chemical Reduction Method (화학적환원에 의한 DNA-mediated 금 나노입자의 합성 및 특성)

  • Sohn, Jun Youn;Sohn, Jeong Sun
    • Applied Chemistry for Engineering
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    • v.26 no.4
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    • pp.515-519
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    • 2015
  • Complexes composed of hydrogen tetrachloroaurate (III) trihydrate ($HAuCl_4{\cdot}3H_2O$) and DNA were first formed for the synthesis of gold nanoparticle using a DNA template, which were validated using UV-Vis spectroscopy. The morphology of complexes were also characterized by scanning electron microscopy (SEM). DNA-mediated gold nanoparticles were synthesized by the chemical reduction of DNA-Au(III) complexes using hydrazine ($N_2H_4$) and sodium borohydride ($NaBH_4$) as reducing agents. The effects of reducing agent types and their concentration on the formation of gold nanoparticles were investigated. The results showed that hydarazine was the most effective for the reduction of DNA-Au(III) complex. The DNA-mediated gold nanoparticles were characterized SEM, particle size analyzer (PSA), and transmission electron microscopy (TEM). Gold nanoparticles with 55~80 nm in diameter were formed by the aggregation of smaller gold nanoparticles (~nm), which was confirmed in the DNA matrix.

Directed Alignment of DNA Molecule between the gold electrodes (금 전극위에 DNA 분자의 정렬에 관한 연구)

  • Hwang, Hyun Suk;Kim, Hyung Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.8
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    • pp.5586-5590
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    • 2015
  • In this paper, the directed alignment methode of the DNA molecule between the Au electrodes was suggested for the application of nano devices. To fabricate the nano device coated DNA, 2-Aminoethanthiol(AET) was coated on Au electrodes which was formed using photo-lithography process on $SiO_2/Si$ substrates. In general, the AET that was a positive charge with $NH^{3+}$ was strongly combined under the electrostatic interaction with DNA molecule which had to be a negative charge. The DNA molecules could be easily aligned between Au electrodes coated with AET. The structures of the DNA molecules were investigated using AFM(Atomic force microscope), they were changed from single types to bundle according to the AET concentrations.

Nuclear DNA Quantification of Some Ceramialean Algal Spermatia by Fluorescence Microscopic Image Processing and their Nuclear SSU rDNA Sequences

  • Choi, Han-Gu;Lee, Eun-Young;Oh, Yoon-Sik;Kim, Hyung-Seop;Lee, In-Kyu
    • ALGAE
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    • v.19 no.2
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    • pp.79-90
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    • 2004
  • Nuclear DNA contents of spermatia from eight ceramiacean and four dasyacean algae (Ceramiales, Rhodophyta) and microspores from two land plants were estimated by fluorescence microscopic image processing and their nuclear SSU rDNA sequence data were analyzed. In frequency distribution patterns, the DAPI-stained nuclear volume (NV) of spermatia showed two peaks corresponding to 1C and 2C. Nuclear 2C DNA contents estimated from NV were 0.45-2.31 pg in ceramiacean and 0.40-0.57 pg in dasyacean algae and 8.42-9.51 pg in two land plants, Capsicum annuum and Nicotiana tabacum. By nuclear patterning of vegetative cells derived from an apical cell, 2C DNA contents of spermatia were 2.31 pg in an alga having uninucleate and non-polyploid nucleus (Aglaothamnion callophyllidicola), 0.45-1.94 pg in algae having uninucleate and polyploid nucleus (Antithamnion spp. and Pterothamnion yezoense), and 0.40-0.62 pg in algae having multinucleate and non-polyploid nuclei (Griffithsia japonica and dasyacean algae). Each mature spermatium and microspore (pollen grain) seemed to have a 2C nucleus, which may provide a genetic buffering system to protect the genetic content of a spermatium and microspore from potentially lethal mutations. Nuclear DNA content and SSU rDNA sequence of Antithamnion sparsum from Korea were reasonably different from those of Antithamnion densum from France. The data did not support the previous taxonomic studies that these two taxa could be conspecific.

In vitro Evidence that Purified Yeast Rad27 and Dna2 are not Stably Associated with Each Other Suggests that an Additional Protein(s) is Required for a Complex Formation

  • Bae, Sung-Ho;Seo, Yeon-Soo
    • BMB Reports
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    • v.33 no.2
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    • pp.155-161
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    • 2000
  • The saccharomyces cerevisiae Rad27, a structure-specific endonuclease for the okazaski fragment maturation has been known to interact genetically and biochemically with Dna2, an essential enzyme for DNA replication. In an attempt to define the significance of the interaction between the two enzymes, we expressed and purified both Dna2 and Rad27 proteins. In this report, Rad27 could not form a complex with Dna2 in the three different analyses. The analyses included glycerol gradient sedimentation, protein-column chromatography, and coinfection of baculoviruses followed by affinity purification. This is in striking contrast to the previous results that used crude extracts. These results suggest that the interaction between the two proteins is not sufficiently stable or indirect, and thus requires an additional protein(s) in order for Rad27 and Dna2 to form a stable physical complex. This result is consistent with our genetic findings that Schizosaccharomyces pombe Dna2 is capable of interacting with several proteins that include two subunits of polymerase $\delta$, DNA ligase I, as well as Fen-1. In addition, we found that the N-terminal modification of Rad27 abolished its enzymatic activity. Thus, as suspected, we found that on the basis of the structure determination, N-terminal methionine indeed plays an important role in the nucleolytic cleavage reaction.

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Funcyional Studies on Gene 2.5 Protein of Bacteriophage T7 : Protein Interactions of Replicative Proteins (박테리오파아지 T7 의 기능에 관한 연구;복제단백질간의 단백질 상호작용)

  • 김학준;김영태
    • Journal of Life Science
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    • v.6 no.3
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    • pp.185-192
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    • 1996
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, is required for T7 DNA replication, recombination, and repair. T7 gene 2.5 protein has two distinctive domains, DNA binding and C-terminal domain, directly involved in protein-protein interaction. Gene 2.5 protein participates in the DNA replication of Bacteriophage T7, which makes this protein essential for the T7 growth and DNA replication. What gene 2.5 protein makes important at T7 growth and DNA replication is its binding affinity to single-stranded DNA and the protein-protein important at T7 DNA replication proteins which are essential for the T7 DNA synthesis. We have constructed pGST2.5(WT) encoding the wild-type gene 2.5 protein and pGST2.5$\Delta $21C lacking C-terminal 21 amino acid residues. The purified GST-fusion proteins, GST2.5(WT) and GST2.5(WT)$\Delta$21C, were used for whether the carboxyl-terminal domain participates in the protein-protein interactions or not. GST2.5(WT) and GST2.5$\Delta$21C showed the difference in the protein-protein interaction. GST2.5(WT) interacted with T7 DNA polymerase and gene 4 protein, but GST2.5$\Delta$21C did not interact with either protein. Secondly, GST2.5(WT) interacts with gene 4 proteins (helicase/primase) but not GST2.5$\Delta$21C. these results proved the involvement of the carboxyl-terminal domain of gene 2.5 protein in the protein-protein interaction. We clearly conclude that carboxy-terminal domain of gene 2.5 protein is firmly involved in protein-protein interactions in T7 replication proteins.

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Ancestral Remnants in the Deoxyribonucleic Acid from Pseudomonas (Deoxyribonucleic Acid 속의 Pseudomonas로부터의 Ancestral Remnants)

  • J. De Ley;In Won Park
    • Journal of the Korean Chemical Society
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    • v.13 no.4
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    • pp.355-364
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    • 1969
  • Cross-hybridizations between DNA of two pseudomonads and a xanthomonad suggested that the three DNA types had a considerable section in common. The existence of this common part was proved by hybridization of preselected DNA, i.e. DNA resulting from a previous hybridization between any one set of two DNA types, with the third type. It was thus shown that about 50% of the DNA of the three organisms was similar. This common part was isolated in pure state and its % (G+C) was found to be indentical to the overall base composition of the native DNA. The evolutionary drift in % (G+C) could thus not be detected. The total molecular weight of the chromosornal DNA/bacterial nucleoid was determined to be 2.4 ${\times} 10^9$daltons. It can therefore be estimated that the common putida-fluorescenspelargonii DNA part consists of some 2,000 cistrons. P. putida and P. fluorescens share an additional 1,300 cistrons, and all xanthomonads share at least an additional 1,000 cistrons.

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A DNA Sequence Generation Algorithm for Traveling Salesman Problem using DNA Computing with Evolution Model (DNA 컴퓨팅과 진화 모델을 이용하여 Traveling Salesman Problem를 해결하기 위한 DNA 서열 생성 알고리즘)

  • Kim, Eun-Gyeong;Lee, Sang-Yong
    • Journal of the Korean Institute of Intelligent Systems
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    • v.16 no.2
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    • pp.222-227
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    • 2006
  • Recently the research for Traveling Salesman Problem (TSP) using DNA computing with massive parallelism has been. However, there were difficulties in real biological experiments because the conventional method didn't reflect the precise characteristics of DNA when it express graph. Therefore, we need DNA sequence generation algorithm which can reflect DNA features and reduce biological experiment error. In this paper we proposed a DNA sequence generation algorithm that applied DNA coding method of evolution model to DNA computing. The algorithm was applied to TSP, and compared with a simple genetic algorithm. As a result, the algorithm could generate good sequences which minimize error and reduce the biologic experiment error rate.

Cellular DNA Repair of Oxidative Deoxyribose Damage by Mammalian Long-Patch Base Excision Repair

  • Sung Jung-Suk;Son Mi-Young
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.103-108
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    • 2005
  • 2-Deoxyribonolactone (dL) arises as a major DNA damage induced by a variety of agents, involving free radical attack and oxidation of C1'-deoxyribose in DNA. We investigated whether dL lesions can be repaired in mammalian cells and the mechanisms underlying the role of DNA polymerase $\beta$ in processing of dL lesions. Pol $\beta$ appeared to be trapped by dL residues, resulting in stable DNA-protein cross-links. However, repair DNA synthesis at site-specific dL sites occurred effectively in cell-free extracts, but predominantly accompanied by long-patch base excision repair (BER) pathway. Reconstitution of long-patch BER demonstrated that FEN1 was capable of removing the displaced flap DNA containing a 5'-dL residue. Cellular repair of dL lesions was largely dependent on the DNA polymerase activity of Pol $\beta$. Our observations reveal repair mechanisms of dL and define how mammalian cells prevent cytotoxic effects of oxidative DNA lesions that may threaten the genetic integrity of DNA.

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Validation of Reduced-volume Reaction in the PowerQuant® System for human DNA Quantification

  • Kim, Hyojeong;Cho, Yoonjung;Kim, Jeongyong;Lee, Ja Hyun;Kim, Hyo Sook;Kim, Eungsoo
    • Biomedical Science Letters
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    • v.26 no.4
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    • pp.275-287
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    • 2020
  • Since its introduction in the forensic field, quantitative PCR (qPCR) has played an essential role in DNA analysis. Quality of DNA should be evaluated before short tandem repeat (STR) profiling to obtain reliable results and reduce unnecessary costs. To this end, various human DNA quantification kits have been developed. Among these kits, the PowerQunat® System was designed not only to determine the total amount of human DNA and human male DNA from a forensic evidence item, but also to offer data about degradation of DNA samples. However, a crucial limitation of the PowerQunat® System is its high cost. Therefore, to minimize the cost of DNA quantification, we evaluated kit performance using a reduced volume of reagents (1/2-volume) using DNA samples of varying types and concentrations. Our results demonstrated that the low-volume method has almost comparable performance to the manufacturer's method for human DNA quantification, human male DNA quantification, and DNA degradation index. Furthermore, using a reduced volume of regents, it is possible to run 2 times more reactions per kit. We expect the proposed low-volume method to cut costs in half for laboratories dealing with large numbers of DNA samples.