• 제목/요약/키워드: DNA concentration

검색결과 1,171건 처리시간 0.033초

Improvement of the electro-transformed cell yield for Bifidobacterium sp. with large DNA

  • Park, Min Ju;Park, Myeong Soo;Ji, Geun Eog
    • 한국식품과학회지
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    • 제51권2호
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    • pp.188-191
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    • 2019
  • In this study, the poor electro-transformant yield of large DNA in bifidobacteria was improved by increasing the DNA concentration, which was amplified by enhancing electroporation conditions: treating the cell wall weakening agent and cell membrane permeabilizing molecule as well as changing the electrical parameter. In the enhanced conditions, the electroporation frequency increased from 15 to 71 times compared to the initial conditions at the same DNA concentration. As the DNA concentration increased, the difference in the electroporation frequency between the two conditions became greater, and the curve of the enhanced conditions seemed to be saturated with a DNA concentration over $4{\mu}g$. The present study provided a clue to the recovery of the electroporation frequency with large DNA and formulated the relationship between the DNA concentration, the DNA size and the electroporation frequency in bifidobacteria. Therefore, this study will contribute to the expansion of molecular genetic studies of bifidobacteria.

Formation Rate of DNA Nanowires According to the APTES Concentration

  • Kim, Taek-Woon;Kim, Nam-Hoon;Roh, Yong-Han
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2008년도 하계학술대회 논문집 Vol.9
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    • pp.143-143
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    • 2008
  • Nanowires are promising options for building nanoscale electronic structures coming from high conductivity of nanowires. In particular, Deoxyribonucleic acid (DNA), which is structurally nanowire, can obtain highly ordered electronic components for nanocircuitry and/or nanodevices because of its very flexible length controllability, nanometer-size diameter, about 2 nm, and self-assembling properties. In this work, we used the method to form DNA-Nanowires (NWs) by using chemical treatment on Silicon (Si) surface, and Aminopropyl-triethoxysilane (APTES) was used as inducer of DNA sequence to modify the characteristics of Si surface. Moreover, we performed tilting technique to align DNA by the direction of flow of DNA solution. We investigated the assembly process between DNA molecules and APTES - coated Si surface according to the APTES concentration, from $1.2{\mu}\ell$ to $120{\mu}\ell$. Atomic Force Microscopy (AFM) images showed the combination rate of DNA molecules by the change of APTES concentration. As APTES concentration becomes thicker, aggregation of DNA molecules occurs, and this makes a kind of DNA networks. In this respect, we confirmed that there's a positive relationship between the concentration of APTES and the formation rate of DNA nanowires. Since there have been lots of research preceded to utilize DNA nanowires as template, so by using this positive relationship with proper alignment technique, realization of nano electronic devices with DNA nanowires might be feasible.

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Contribution of Counterion Entropy to the Salt-Induced Transition Between B-DNA and Z-DNA

  • Lee, Youn-Kyoung;Lee, Juyong;Choi, Jung Hyun;Seok, Chaok
    • Bulletin of the Korean Chemical Society
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    • 제33권11호
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    • pp.3719-3726
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    • 2012
  • Formation of Z-DNA, a left-handed double helix, from B-DNA, the canonical right-handed double helix, occurs during important biological processes such as gene expression and DNA transcription. Such B-Z transitions can also be induced by high salt concentration in vitro, but the changes in the relative stability of B-DNA and Z-DNA with salt concentration have not been fully explained despite numerous attempts. For example, electrostatic effects alone could not account for salt-induced B-Z transitions in previous studies. In this paper, we propose that the B-Z transition can be explained if counterion entropy is considered along with the electrostatic interactions. This can be achieved by conducting all-atom, explicit-solvent MD simulations followed by MM-PBSA and molecular DFT calculations. Our MD simulations show that counterions tend to bind at specific sites in B-DNA and Z-DNA, and that more ions cluster near Z-DNA than near B-DNA. Moreover, the difference in counterion ordering near B-DNA and Z-DNA is larger at a low salt concentration than at a high concentration. The results imply that the exclusion of counterions by Z-DNA-binding proteins may facilitate Z-DNA formation under physiological conditions.

Synthesis and characterization of doxorubicin hydrochloride drug molecule-intercalated DNA nanostructures

  • Gnapareddy, Bramaramba;Deore, Pragati Madhukar;Dugasani, Sreekantha Reddy;Kim, Seungjae;Park, Sung Ha
    • Current Applied Physics
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    • 제18권11호
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    • pp.1294-1299
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    • 2018
  • In this paper, we demonstrate the feasibility of constructing DNA nanostructures (i.e. DNA rings and double-crossover (DX) DNA lattices) with appropriate doxorubicin hydrochloride (DOX) concentration and reveal significant characteristics for specific applications, especially in the fields of biophysics, biochemistry and medicine. DOX-intercalated DNA rings and DX DNA lattices are fabricated on a given substrate using the substrateassisted growth method. For both DNA rings and DX DNA lattices, phase transitions from crystalline to amorphous, observed using atomic force microscopy (AFM) occurred above a certain concentration of DOX (at a critical concentration of DOX, $30{\mu}M$ of $[DOX]_C$) at a fixed DNA concentration. Additionally, the coverage percentage of DNA nanostructures on a given substrate is discussed in order to understand the crystal growth mechanism during the course of annealing. Lastly, we address the significance of optical absorption and photoluminescence characteristics for determining the appropriate DOX binding to DNA molecules and the energy transfer between DOX and DNA, respectively. Both measurements provide evidence of DOX doping and $[DOX]_C$ in DNA nanostructures.

Relaxation Process of the Photoexcited State and Singlet Oxygen Generating Activity of Water-soluble meso-Phenanthrylporphyrin in a DNA Microenvironment

  • Hirakawa, Kazutaka;Ito, Yusuke;Yamada, Takashi;Okazaki, Shigetoshi
    • Rapid Communication in Photoscience
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    • 제3권4호
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    • pp.81-84
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    • 2014
  • To examine the microenvironmental effect of DNA on the photosensitized reaction, the electron-donor-connecting porphyrin, meso-(9-phenanthryl)-tris(N-methyl-p-pyridinio) porphyrin (Phen-TMPyP), was synthesized. Phen-TMPyP can bind to oligonucleotides with two binding modes, depending on the DNA concentration. The fluorescence lifetime measurement of Phen-TMPyP shows a shorter component than that of the reference porphyrin without the phenanthryl moiety. However, the observed value is much longer than those of previously reported similar types of electron-donor-connecting porphyrins, suggesting that electron-transfer quenching by the phenanthryl moiety is not sufficient. The fluorescence quantum yield of Phen-TMPyP ($5{\mu}M$) decreased with an increase in DNA concentration of up to $5{\mu}M$ base pair (bp), possibly due to self-quenching through an aggregation along the DNA strand, increased with an increase in DNA concentration of more than $5{\mu}M$ bp and reached a plateau. The fluorescence quantum yield of Phen-TMPyP with a sufficient concentration of DNA was larger than that of the reference porphyrin. The singlet oxygen ($^1O_2$) generating activity of Phen-TMPyP was confirmed by the near-infrared emission spectrum measurement. The quantum yield of $^1O_2$ generation was decreased by a relatively small concentration of DNA, possibly due to the aggregation of Phen-TMPyP, and recovered with a sufficient concentration of DNA. The recovered quantum yield was rather smaller than that without DNA, indicating the quenching of $^1O_2$ by DNA. These results show that a DNA strand can stabilize the photoexcited state of a photosensitizer and, in a certain case, suppresses the $^1O_2$ generation.

Estimation of the Efficiency of Transgenic Rabbit Production Following GFP Gene Microinjection into Rabbit Zygotes

  • Jin, D.I.;Im, K.S.;Kim, D.K.;Choi, W.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권10호
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    • pp.1367-1372
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    • 2000
  • The present study was conducted to evaluate the efficiency of transgenic rabbit production by DNA microinjection using EGFP (Enhanced Green Fluorescent Protein) gene. In this experiment EGFP coding sequences fused to CMV promoter were microinjected into rabbit one-cell embryos, and then GFP expression and gene integration were evaluated in preimplantation embryos and fetuses recovered on day 15 of pregnancy to determine efficiency of transgenic rabbit production. Effect of DNA concentration was also tested on development in vitro following microinjection and transgene integration in fetuses. Development of embryos in vitro was decreased by DNA microinjection, but the rates of pregnancy and implantation were not significantly affected by microinjection. As development progressed in vitro percentage of GFP expression in rabbit embryos was decreased, resulting GFP expression detected in 37.5% of blastocysts. The efficiencies for production of transgenic fetuses were 4.0% and 7.6%, respectively, when $10ng/{\mu}l$ and $20ng/{\mu}l$ of DNA concentration were microinjected. Transgenic fetuses were confirmed by GFP expression and PCR analysis of fetus genomic DNA. These results indicated that DNA microinjection itself damaged embryo development and DNA concentration affected the efficiency of transgenic rabbit production.

Investigation of Circulating Cell-Free DNA Concentration in Dogs with Pancreatitis

  • Jae-Hun Kim;Soo-Yeon Jeong;Chul Park
    • 한국임상수의학회지
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    • 제41권3호
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    • pp.143-149
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    • 2024
  • Circulating cell-free DNA (cfDNA) constitutes a fragment of DNA released into the blood through cellular apoptosis or necrosis. In human medicine, cfDNA has been studied as a disease severity biomarker. Recent studies have shown that concentrations of cfDNA in dogs with immune-mediated and tumor-related diseases are increased. Pancreatitis is known to be caused by excessive release of trypsin, which leads to edema, inflammation, necrosis, and apoptosis in the pancreas. Based on the results of research showing an increase of cfDNA due to apoptosis and necrosis of cells, we hypothesized that cfDNA concentration would increase in the presence of pancreatitis. A total of 35 dogs were studied, including 21 with pancreatitis and 14 without any inflammatory diseases (normal group). The results showed that the concentration of cfDNA in dogs with pancreatitis was approximately twice that of normal dogs (median 0.0912 ng/μL. p-value 0.028). This result suggests that cfDNA can serve as a new biomarker for estimating pancreatitis severity.

M-DNA 전기 특성의 금속 이온 농도 및 합성 온도 의존성 (Dependence of Electrical Properties of M-DNA on Metal Ion Concentration and Synthetic Temperature)

  • 유환일;김경섭;김남훈;노용한
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2007년도 추계학술대회 논문집
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    • pp.105-106
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    • 2007
  • M-DNAs based on zinc and cobalt ions were formed at the conditions of $26^{\circ}C$, 76% humidity, atmosphere, and pH 8.0. Some process parameters in synthesis of M-DNA such as synthetic temperature, concentration of metal ions, and synthetic time were varied and the electrical properties were investigated by changes of the parameters. The electrical properties of M-DNA showed the dependences on synthetic temperature, concentration of metal ions and synthetic time. The I-V characteristics rapidly increased with each increase of the parameters.

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total DNA 및 단백질 함량변화에 의한 C. polykrikoides 조기적조 예측 응용 (Application of DNA Content and Total Protein Concentration to Predict Blooms Caused by Cochlodinium polykrikoides (Dinophyceae) in Korean Coastal Waters)

  • Cho, Eun-Seob;Park, Yong-Kyu
    • 생명과학회지
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    • 제14권2호
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    • pp.255-262
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    • 2004
  • 본 연구는 남해안에서 발생되는 유해성 C. polykrikoides 조기적조를 예측하기 위한 기법으로 DAPI로 염색시킨 DNA와 단백질 함량변화를 단기간으로 조사했다. 조사기간 중의 환경요인, 영양염 (질산, 아질산, 인산염) 농도는 조사지점이나 시기에 관계없이 거의 비슷하게 보였다. 그러나 C. polykrikoides 밀도는 조사지점에 따라 현저하게 다르게 나타났다. 2000년 8월 초순의 경우 C2, C5, C6에서 리터 당34, 62, 57세포를 각각 출현했으며, 8월 중순에는 C3에서 최고 547 세포가 보였다. C. polykrikoides 출현밀도와 DNA 및 단백질 함량과는 양성적인 상관관계를 보였다. 특히 C. polykrikoides 세포밀도가 아주 낮을 경우에 높은 상관값을 나타내었다. 따라서 DNA 및 단백질 함량변화 기법은 C. polykrikoides 조기적조를 쉽게 예측 할 수 있는 중요한 도구도 이용될 수 있다.

SV 40 바이러스가 유도한 DNA 합성효소의 특성에 대한 연구 (Characterizations of DNA-polymerases Induced by SV40 Virus Infection of African Green Monkey Kidney Cells (AGMK))

  • 강현삼
    • 미생물학회지
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    • 제14권3호
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    • pp.135-145
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    • 1976
  • Confluent AGMK cells were infected by large plaque SV40 virus. Levels of DNA polymeras $({\alpha}\;and\;{\beta})$ were measured in the cytoplasm and the cell nucleus. The activities of DNA $polymerase-{\alpha}$ which found in both the cell nucleus and the cytoplasm were increased approximately eight folds at 48 hours after infection of SV40 virus. Only insignificant but constant amounts of DNA $polymerase-{\beta}$ were found either in the nucleus of the SV40 infected cell or of the uninfected cell. The characteristics of the SV40 virus induced DNA polymerases were compared with that of the uninfected cellular DNA polymerase in regard of the effects of pH, salt concentration, NEM concentration and temperature on those enzyme activities. No differential effect was found between both enzymes. Endouclease activities wre examined in the purified DNA $polymerase-{\alpha}\;and\;{\beta}$. The low level of endonuclease activity which might cut SV40 DNA 1 at one site was observed in the DNA $polymerase-{\alpha}$ whereas high but nonspecific endonuclease activities were found in the DNA $polymerase-{\beta}$.

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