• Title/Summary/Keyword: DNA separation

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NCAPH Stabilizes GEN1 in Chromatin to Resolve Ultra-Fine DNA Bridges and Maintain Chromosome Stability

  • Kim, Jae Hyeong;Youn, Yuna;Hwang, Jin-Hyeok
    • Molecules and Cells
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    • v.45 no.11
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    • pp.792-805
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    • 2022
  • Repairing damaged DNA and removing all physical connections between sister chromosomes is important to ensure proper chromosomal segregation by contributing to chromosomal stability. Here, we show that the depletion of non-SMC condensin I complex subunit H (NCAPH) exacerbates chromosome segregation errors and cytokinesis failure owing to sister-chromatid intertwinement, which is distinct from the ultra-fine DNA bridges induced by DNA inter-strand crosslinks (DNA-ICLs). Importantly, we identified an interaction between NCAPH and GEN1 in the chromatin involving binding at the N-terminus of NCAPH. DNA-ICL activation, using ICL-inducing agents, increased the expression and interaction between NCAPH and GEN1 in the soluble nuclear and chromatin, indicating that the NCAPH-GEN1 interaction participates in repairing DNA damage. Moreover, NCAPH stabilizes GEN1 within chromatin at the G2/M-phase and is associated with DNA-ICL-induced damage repair. Therefore, NCAPH resolves DNA-ICL-induced ultra-fine DNA bridges by stabilizing GEN1 and ensures proper chromosome separation and chromosome structural stability.

Simple Screening Method for Double-strand DNA Binders Using Hairpin DNA-modified Magnetic Beads

  • Jo, Hun-Ho;Min, Kyoung-In;Song, Kyung-Mi;Ku, Ja-Kang;Han, Min-Su;Ban, Chang-Ill
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.247-250
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    • 2011
  • We designed an effective screening method for double strand DNA (dsDNA) binders using DNA-modified magnetic particles. Hairpin DNA was immobilized on the surface of magnetic particle for a simple screening of dsDNA binding materials in a solution containing various compounds. Through several magnetic separation and incubation processes, four DNA-binding materials, DAPI, 9AA, AQ2A, and DNR, were successfully screened from among five candidates. Efficiency of screening was demonstrated by HPLC analysis using a C2/18 reverse-phase column. In addition, their relative binding strengths were verified by measuring the melting temperature ($T_m$). If hairpin DNA sequence is modified for other uses, this magnetic bead-based approach can be applied as a high-throughput screening method for various functional materials such as anti-cancer drugs.

Detection of Fish Virus by Using Immunomagnetic Separation and Polymerase Chain Reaction (IMS-PCR)

  • KIM Soo Jin;OH Hae Keun;CHOI Tae-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.6
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    • pp.948-955
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    • 1997
  • Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

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Megabase-sized DNA isolation and electrophoretic karyotype of fusarium oxysporum schlecht

  • Park, Min-Seon;Min, Byung-Re
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.132-135
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    • 1995
  • To investigate the electrophoretic karytype of Fusarium oxysporum, intact chromosomal DNA was separated by pulsed-field gel electrophoresis (PEGE). DNA extraction from nulcei, mycelia and protoplasts were compared with one another and with the quantity and the suitability for PFGE separation in agarose gel. As a result, the most useful extracting method for intact DNA was found to be that from protoplasts. By varying the electrophoretic conditions, 8 chromosomal DNA bounds were resolved. Using the Schizosaccharomyces pombe and Saccharomyces cerevisiae as size standards, the size of Fusarium oxysporum chromosomes was estimated to range from approximately 0.6 Mb TO 6.7 Mb, and total genome size was 26.7 Mb. The suitability of electrophoretic karyotyping as a tool for strain characterization is discussed.

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Genetic Synthesis and Applications of Repetitive Protein Polymers (반복단위 단백질 고분자의 유전공학적 합성 및 응용)

  • Park, Mi-Sung;Choi, Cha-Yong;Won, Jong-In
    • KSBB Journal
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    • v.22 no.4
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    • pp.179-184
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    • 2007
  • This study introduces the characteristics and some applications of repetitive polypeptides, especially to the biomaterial, tissue engineering scaffolds, drug delivery system, and DNA separation systems. Since some fibrous proteins, which consist of repeating peptide monomers, have been reported that their physical properties are changed dramatically by means of temperature alteration or pH shifting. For that reason, fibrous protein-mimetic polypeptides, which are produced by the recombinant technology, can be applied to the diverse biological fields. Repetitive polypeptides can also be used in the bioseparation area such as DNA sequencing, because they make DNA separation possible in free-solution electrophoresis by conjugating DNA fragments to them. Moreover, artificial synthesis of repetitive polypeptides helps to demonstrate the correlations between mechanical properties and structures of natural protein polymer, which have been proven that repetitive domains are affected by the sequence of the repeating domains and the number of repeating subunits. Repetitive polypeptides can be biologically synthesized using some special cloning methods, which are represented here. Recursive directional ligation (RDL) and controlled cloning method (CCM) have been proposed as excellent cloning methods in that we can control the number of repetition in the multimerization of polypeptides and the components of repetitive polypeptides by either method.

Network Address Translation By Flow Separation (데이터 플로우 구별에 의한 네트워크 주소 변환)

  • 윤승용;이광희;최창국;전우직
    • Proceedings of the Korean Information Science Society Conference
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    • 2000.10c
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    • pp.393-395
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    • 2000
  • 현재 인터넷이 직면하고 있는 IP 주소 부족문제 해결을 위한 새로운 방안으로서 데이터 플로우 구별에 의한 네트워크 주소 변환(NAT-FS : NAT by Flow Separation) 기법을 제안하고 기술한다. 이 방식은 기존의 NAPT와 같이 단 하나의 글로벌 IP 주소에 모든 로컬 호스트가 할당되면서도 Basic NAT 방식처럼 DNA와 연동하여 Full Access 기능도 지원할 수 있다.

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Code Optimization in DNA Computing for the Hamiltonian Path Problem (해밀톤 경로 문제를 위한 DNA 컴퓨팅에서 코드 최적화)

  • 김은경;이상용
    • Journal of KIISE:Software and Applications
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    • v.31 no.4
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    • pp.387-393
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    • 2004
  • DNA computing is technology that applies immense parallel castle of living body molecules into information processing technology, and has used to solve NP-complete problems. However, there are problems which do not look for solutions and take much time when only DNA computing technology solves NP-complete problems. In this paper we proposed an algorithm called ACO(Algorithm for Code Optimization) that can efficiently express DNA sequence and create good codes through composition and separation processes as many as the numbers of reaction by DNA coding method. Also, we applied ACO to Hamiltonian path problem of NP-complete problems. As a result, ACO could express DNA codes of variable lengths more efficiently than Adleman's DNA computing algorithm could. In addition, compared to Adleman's DNA computing algorithm, ACO could reduce search time and biological error rate by 50% and could search for accurate paths in a short time.

Polyamine Group Assembled Silica Coated Ferrite Nanoparticle for Lambda DNA Detection

  • Park, Moo-Eon;Chang, Jeong-Ho
    • Proceedings of the Korean Powder Metallurgy Institute Conference
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    • 2006.09b
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    • pp.1193-1194
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    • 2006
  • The magnetic ferrite nanoparticles were synthesized and coated by silica precursor in controlling the coating thicknesses and sizeses. The surface modification was performed with amino-functionalized organic silanes on silica coated magnetic nanoparticles. The use of functionalized self-assembled magnetic ferrite nanoparticles for nucleic acid separation process give a lot of advantages rather than the conventional silica based process.

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PCR of Gut Contents for a Food Web Study of a Marine Ecosystem

  • Kim, Nack-Keun;Kim, Kyoung-Sun;Kim, Hyun-Woo
    • Fisheries and Aquatic Sciences
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    • v.10 no.4
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    • pp.179-185
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    • 2007
  • Understanding dietary habits is one of the most important factors in studying food webs and other ecological processes. Here we designed universal primers to amplify portions of the 18S and 28S rDNA sequences to examine gut contents using PCR techniques. The gut contents of sailfin sandfish (Arctoscopus japonicus) and pacific squid (Todarodes pacificus) were examined. In total, 11 families of prey were identified with 18S and 28S rDNA using the universal primers. The DNA sequence data indicated that the primer sets successfully amplified a wide spectrum of species and represented gut contents in a relatively convenient way. We found that information in the NCBI database was not yet sufficient to discriminate the species we isolated. In addition, technology for the separation of heterogeneous PCR products and better resolution and quantification protocols would help increase data accuracy.