• 제목/요약/키워드: double-stranded DNA

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Foldback Intercoil DNA and the Mechanism of DNA Transposition

  • Kim, Byung-Dong
    • Genomics & Informatics
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    • 제12권3호
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    • pp.80-86
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    • 2014
  • Foldback intercoil (FBI) DNA is formed by the folding back at one point of a non-helical parallel track of double-stranded DNA at as sharp as $180^{\circ}$ and the intertwining of two double helixes within each other's major groove to form an intercoil with a diameter of 2.2 nm. FBI DNA has been suggested to mediate intra-molecular homologous recombination of a deletion and inversion. Inter-molecular homologous recombination, known as site-specific insertion, on the other hand, is mediated by the direct perpendicular approach of the FBI DNA tip, as the attP site, onto the target DNA, as the attB site. Transposition of DNA transposons involves the pairing of terminal inverted repeats and 5-7-bp tandem target duplication. FBI DNA configuration effectively explains simple as well as replicative transposition, along with the involvement of an enhancer element. The majority of diverse retrotransposable elements that employ a target site duplication mechanism is also suggested to follow the FBI DNA-mediated perpendicular insertion of the paired intercoil ends by non-homologous end-joining, together with gap filling. A genome-wide perspective of transposable elements in light of FBI DNA is discussed.

버섯에 존재하는 다양한 dsRNA의 분자생물학적 연구 (Molecular characteristics of diverse dsRNAs in edible fungi)

  • 박윤정;신평균;장갑열;공원식;정종천;유영복
    • 한국버섯학회지
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    • 제8권2호
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    • pp.41-50
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    • 2010
  • Mycoviruses have been found in many fungal species including mushrooms. Double-stranded (ds) RNA genomes were common type in mycoviruses, but single-stranded (ss) RNA mycoviruses were also reported in some fungal species. Sequencing analysis using cDNA cloning experiments revealed that mycoviruses can be classified into several different virus families such as Totiviridae, Hypoviridae, Partitiviridae and Barnaviridae etc. Because the nucleotide sequence data that are available in these days are very limited in a number of mycoviruses, the existence of more diverse viral groups in fungi are currently expected. In this review, we selected four different fungal groups, which were considered as the model systems for mycovirus related studies in both plant pathogenic fungi and edible mushroom species, and discussed about their molecular characteristics of diverse mycoviruses. The plant pathogenic fungi introduced here were Cryphonectria parasitica and Helminthosporium victoriae and the edible mushroom species were Agaricus bisporus and Pleurotus ostreatus.

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Characterization of denaturation and renaturation of DNA for DNA hybridization

  • Wang, Xiaofang;Lim, Hyun Jeong;Son, Ahjeong
    • Environmental Analysis Health and Toxicology
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    • 제29권
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    • pp.7.1-7.8
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    • 2014
  • Objectives The present study was designed to systematically characterize the denaturation and the renaturation of double stranded DNA (dsDNA), which is suitable for DNA hybridization. Methods A series of physical and chemical denaturation methods were implemented on well-defined 86-bp dsDNA fragment. The degree of each denaturation was measured and the most suitable denaturation method was determined. DNA renaturation tendency was also investigated for the suggested denaturation method. Results Heating, beads mill, and sonication bath did not show any denaturation for 30 minutes. However probe sonication fully denatured DNA in 5 minutes. 1 mol/L sodium hydroxide (alkaline treatment) and 60% dimethyl sulfoxide (DMSO) treatment fully denatured DNA in 2-5 minutes. Conclusions Among all the physical methods applied, the direct probe sonication was the most effective way to denature the DNA fragments. Among chemical methods, 60% DMSO was the most adequate denaturation method since it does not cause full renaturation during DNA hybridization.

전계효과 트랜지스터(FETs)를 이용한 전하 검출형 DNA 센서에서 Debye length에 따른 검출 감도 (Sensitivity of a charge-detecting label-free DNA sensor using field-effect transistors (FETs) depending on the Debye length)

  • 송광섭
    • 전자공학회논문지SC
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    • 제48권2호
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    • pp.86-90
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    • 2011
  • 전계효과 트랜지스터(FETs)를 이용한 전하 검출형 DNA센서는 DNA가 가지고 있는 음전하를 중성화 시키는 양이온의 영향은 매우 중요하다. 본 논문에서는 양이온 농도에 의존하는 Debye length에 관한 연구를 통해 DNA 검출감도를 평가하였다. Debye length는 낮은 농도의 NaCl 용액에서 긴 거리를 유지하며, Debye length가 높은 용액에서 DNA가 가지고 있은 음전하는 게이트 채널에 보다 많은 영향을 미친다. 용액내 NaCl농도가 1 mM인 버퍼 용액에서 상보적 DNA의 hybridization에 의한 전계효과 트랜지스터의 게이전압은 21 mV 시프트 했으며, NaCl 농도가 10 mM인 버퍼 용액에서는 7.2 mV, NaCl농도가 100 mM인 버퍼 용액에서는 전계효과 트랜지스터의 게이트 전압이 5.1 mV 각각 시프트 하였다. 이러한 결과를 바탕으로 전계효과 트랜지스터를 이용한 전하 검출형 DNA센서의 검출 감도는 Debye length에 의존하는 것을 규명하였다.

미소전극어레이형 DNA칩을 이용한 유전자다형의 전기화학적 검출 (Electrochemical Detection of Single Nucleotide Polymorphism (SNP) Using Microelectrode Array on a DNA Chip)

  • 최용성;권영수;박대희
    • 대한전기학회논문지:전기물성ㆍ응용부문C
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    • 제53권5호
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    • pp.286-292
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    • 2004
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 ㎷/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic System.

Analysis of Double-Stranded DNA Fragments by Capillary Electrophoresis Using Entangle Polymer Solutions in Uncoated Fused Silica Capillary Columns

  • Lee, Jong-Jin;Lee, Kong-Joo
    • BMB Reports
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    • 제31권4호
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    • pp.384-390
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    • 1998
  • DNA fragments (51-587 bp) were separated by capillary electrophoresis using entangled polymer, hydroxyethylcellulose, in uncoated fused silica capillary columns. The factors affecting the separation of DNA fragments with hydroxyethylcellulose media were evaluated, i.e., the concentration of buffer and entangled polymer, effects of additives (methanol, ethidium bromide, EDTA), temperature, and injection methods. Maximum performance was obtained by adding 5% methanol in 0.5% hydroxyethylcellulose solution at $30^{\circ}C$. Addition of methanol in polymer media increased the resolution of small size DNA fragments (< 100 bp). On the other hand, addition of ethidium bromide and EDTA, which are commonly used in conventional DNA separation, reduced the resolution of DNA fragments in the polymer solution. It turns out that the separation behavior of DNA in entangled polymer is more sensitive to the running condition compared to that in polyacrylamide gel-filled capillary, but the reproducibility of DNA separation in entangled polymer is reliable.

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미소전극형 DNA칩 어레이를 이용한 유전자의 검출 (A Study on Electrical Properties of Dendrimer)

  • 최용성;이경섭
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
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    • pp.1324-1326
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    • 2006
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

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고대 유전자에 대한 두 종류의 DNA 분리 방법의 비교 연구: 실리카 현탁액 방법 및 초원심분리 농축 방법 (The comparative study of two extraction methods for ancient DNA: silica suspension method and ultracentrifugal concentrator method)

  • 이은정
    • 분석과학
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    • 제31권2호
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    • pp.65-70
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    • 2018
  • 이 연구에서는 대규모 병렬형 염기서열 분석 (massively parallel sequencing)에 적용할 샷건 라이브러리 (shotgun library)를 성공적으로 제작하기 위해 두 가지 유형의 고대 DNA (ancient DNA, aDNA) 분리 방법을 비교하였다. 헝가리 선사 시대 늑골 뼈 시료로 실리카 현탁액을 이용한 추출법과 Amicon Ultracel-15 10K 초원심 분리 장치(Millipore)를 이용한 추출법을 비교하였다. 약 150 mg의 뼛가루에서 각각의 방법으로 3 회 반복 추출한 후 이중 가닥 DNA (double stranded DNA, ds DNA)의 양을 측정하였다. 초원심분리 농축 방법은 실리카 현탁액을 사용하는 것보다 더 빠르고, 더 쉬운 공정이며 약 11 배 높은 DNA 회수율을 나타냈다. 또한 초원심 분리 장치로 획득한 DNA 주형은 실리카 현탁액으로 획득한 것보다 샷건 라이브러리가 훨씬 성공적으로 만들어졌다. 두 종류의 aDNA 추출 방법을 비교한 본 연구는 Amicon 장치를 사용하는 분리법이 시간의 절약, 단순한 프로세스 및 높은 효율 등의 장점을 지니고 있음을 보여주었다.

한국의 양식대하에서의 흰반점증상 바이러스감염의 특징 (Characterization of the White Spot Syndrome Baculovirus (WSBV) Infection In Fresh Shrimp, Penaeus chinensis, Cultured in Korea)

  • 허문수
    • 생명과학회지
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    • 제15권2호
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    • pp.248-252
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    • 2005
  • 대하 새우양식장의 대하에 흰반점 증상을 나타내는 원인바이러스는 막대형의 이중막을 가지고 있었으며, 전자 현미경 관찰 결과 평균 크기가 $250\~300\times70\;nm$였고, 조직학적 병변은 위상피 등에서 핵이 비대해지는 것이 관찰되었다. 공격실험에서는 건강체의 새우에 많은 누적 폐사율을 보였다. 원인 바이러스 단백질은 21개의 밴드를 보였으며, 핵산 분석 결과 total 분자량은 114 kb로 나타났다.

Functional Analysis of RAD4 Gene Required for Nucleotide Excision Repair of UV-induced DNA Damage in Saccharomyces cerevisiae

  • Park, Sang Dai;Park, In Soon
    • Animal cells and systems
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    • 제6권4호
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    • pp.311-315
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    • 2002
  • The RAD4 gene is essential for nucleotide excision repair in Saccharomyces cerevisiae. It has been known that the deduced amino acid sequence of Rad4 protein contains three DNA-dependent ATPase/helicase motifs. To determine the biochemical activities and functional role of RAD4 the Rad4 protein was expressed and purified. Immunoblot analysis showed a specific band of 21 kDa, which was well-matched with the size of open reading frame of the RAD4 gene. The purified Rad4 protein had no detectable helicase activity. However, the protein could interact with double stranded oligonucleotides, as judged by mobility shift assay. This result suggests that the Rad4 protein is a DNA binding protein.