• Title/Summary/Keyword: DNA Manipulation

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Micromachined DNA Manipulation Device Using Circular Multi-Electrodes (원형 다중전극을 이용한 DNA 조작소자)

  • Moon, Sang-Jun;Yun, Jae-Young;Lee, Seung-S.;Nam, Hong-Kil;Chi, Yeun-Tae
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.27 no.7
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    • pp.1071-1075
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    • 2003
  • In this paper, we present a DNA manipulation device in the reaction chamber, which consists of a center electrode and circular outer electrodes of a reaction unit. The charged bio-molecules, DNA, are manipulated by the charge of the electrode in reaction unit. Controlling the induced dynamic electric field between the center electrode and the outer electrodes, concentration / repulsion / manipulation of bio-molecules are enabled at a periphery of electrode. Concentration of the fluorescent DNA at the center electrode is observed by applying +2V. Subsequently, applying -2V, the concentrated DNA is repelled rapidly from the center electrode, which makes dispersion completely in 0.5second. Furthermore, repeated applying +1V/-1V every 5 seconds at each outer electrode, we can circulate the DNA. We also investigate a micro-heater and sensor for DNA manipulation and reaction temperature. The coefficient of heat-resistance and heater temperature characteristic is 0.0043 and 100$^{\circ}C$/sec, respectively.

Structure-function analysis of PRDI DNA polymerase

  • Jung, Guhung
    • The Microorganisms and Industry
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    • v.16 no.3
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    • pp.6-14
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    • 1990
  • PRDI DNA polymerase is the smallest member of the family B DNA polymerase (Jung et al., 1987). This DNA polyerase is specified by bacteriophage PRDI which infects a wide variety of gram-negative bacteria(Mindich and Bamford, 1988). Because PRDI is highly amenable to genetic and biochemical manipulation, it is a convenient model system with which to study structure-function relationships of DNA polymerase molecules. To determine the functional roles of the highly conserved regions of the family B DNA polymerases, we have initiated site-directed mutagenesis with PRD1 DNA polymerase, and our results show that mutations at the conserved regions within PRD1 DNA polymerase inactivate polymerase complementing activity and catalytic activity.

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A Micromethod for Rapid and Simple Isolation of Genomic DNA from Small Scale Culture of Bifidobacterium (소량의 Bifidobacterium 배양액에서 genomic DNA 추출을 위한 신속/간단한 방법)

  • Jeakal, Soo;Park, Hee-Kyung;Song, Ji-Eun;Heo, Tae-Ryeon;So, Jae-Seong
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.781-783
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    • 1995
  • A method is described for the rapid and simple isolation of genomic DNA from 3 ml culture of Bifidobacterium. The method is expected to be used in gene manipulation of Bifidobacterium spp. The isolated DNA using this method is shown to be an excellent substrate for restriction endonuclease digestion and ligation with T4 DNA ligase.

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Manipulation of Mini-Yeast Artificial Chromosome Containing Xylan Metabolism Related Genes and Mitotic Stability Analysis in Yeast (Xylan 대사유전자를가진미니효모인공염색체의가공및 Mitotic Stability 분석)

  • Da-In Kang;Yeon-Hee Kim
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.436-440
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    • 2022
  • In this study, yeast artificial chromosome Insert (YAC) harboring genes which related xylan metabolism was constructed by using chromosome manipulation technique. For efficient chromosome manipulation, each splitting fragment (DNA module) required for splitting process was prepared and these DNA modules were transformed into Saccharomyces cerevisiae strain YKY164. By two-rounds chromosome splitting, yeast chromosome VII (1,124 kb) was split 887 kb-YAC, 45 kb-mini YAC and 198 kb-YAC and YKY183 strain containing 18 chromosomes was constructed. Splitting efficiency for chromosome manipulation was 50- 78% and expression level of foreign genes on 45 kb-mini YAC and enzyme activity were indistinguishable from that of the YKY164 strain. Furthermore, xylan-degraded products by recombinant enzymes were confirmed and mini-yeast artificial chromosome maintained stable mitotic stability without chromosome loss during 160 generations.

Atomic Force Microscopy(AFM) based Single Cell Manipulation and High Efficient Gene Delivery Technology (원자간력 현미경을 이용한 단일세포 조작 및 고효율 유전자 도입기술)

  • Han, Sung-Woong;Nakamura, Chikashi;Miyake, Jun;Kim, Woo-Sik;Kim, Jong-Min;Chang, Sang-Mok
    • Korean Chemical Engineering Research
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    • v.47 no.5
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    • pp.538-545
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    • 2009
  • The principle and application of a scanning probe microscopy(SPM) are reviewed briefly, and a low-invasive single cell manipulation and a gene delivery technique using an etched atomic force microscopy(AFM) probe tip, which we call a nanoneedle, are explained in detail. The nanoneedle insertion into a cell can be judged by a sudden drop of force in a force-distance curve. The probabilities of nanoneedle insertion into cells were 80~90%, which were higher than those of typical microinjection capillaries. When the diameter of the nanoneedle was smaller than 400 nm, the nanoneedle insertion into a cell over 1 hour had almost no influence on the cell viability. A highly efficient gene delivery and a high ratio of expressed gene per delivered DNA compared the conventional major nonviral gene delivery methods could be achieved using the gene modified nanoneedle.

State-of-the Art Review in Nano-Biomanipulation Technologies (나노-바이오 매니퓰레이션 기술의 현황 및 전망)

  • Kim Deok-Ho;Kim Byungkyu;Park Jong-Oh;Ju Byeong-Kwon
    • Journal of Institute of Control, Robotics and Systems
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    • v.11 no.4
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    • pp.353-362
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    • 2005
  • This article describes a state-of-the art review in nano-biomanipulation technologies. Nanomanipulation of biological objects enables an in-depth study of single molecules such as DNA and RNA, and of biophysical events at the molecular level like molecular motors. Controlled nanomanipulation is challenging but essential for precisely engineering biomolecules or cells and for manufacturing functional nano-biosystems. In this paper, we summarize several contact, non-contact and hybrid methods available for nanomanipulation of biological objects. Advantages currently available methods and their limitations are also compared. Finally, we discuss possible applications of nano-biomanipulation technologies to life science and molecular medicine including cell biology, genetic engineering, biophysics, and biochemistry.

A Rapid Small Scale Method for Extraction of Genomic DNA from Lactobacillus spp. (Lactobacillus spp 로부터 Genomic DNA추출을 위한 신속/간단한 방법)

  • 이석용
    • KSBB Journal
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    • v.15 no.4
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    • pp.411-413
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    • 2000
  • A method is described for the rapid and simple isolation of genomic DNA from 3 mL culture of Lactobacillus crispatus KLB46 The isolated DNA using this method was shown to be an excellent substrate for restriction endonclease digestion and PCR. The method is expected to be used in gentic manipulation of L. crispatus KLB46.

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Design and Control of a New Micro End-effector for Biological Cell Manipulation

  • Shim, Jae-Hong;Cho, Sung-Yong;Cho, Young-Im;Kim, Deok-Ho;Kim, Byung-Kyu
    • 제어로봇시스템학회:학술대회논문집
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    • 2003.10a
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    • pp.2445-2450
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    • 2003
  • Recently, biological technology industry shows great development. Instruments and systems related biological technology have been developed actively. In this paper, we developed a new micro end-effector for biological cell manipulation. The existing micro end-effector for biological cell manipulation has not any force sensing mechanism. Usually, excessive contact force occurring when the end-effector and a cell collide might make a damage on the cell. However, unfortunately, user can not notice the condition in case of using the existing end-effector. In order to overcome we proposed the improved micro end-effector having a force sensing mechanism. This paper presents the design concepts of the new micro end-effector. We carried out calibration of the force sensor and tested the performance of the proposed micro end-effector. Through a series of experiments the new micro end-effector shows the possibility of application for precision biological cell manipulation such as DNA operation

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The Safety of Food Developed by Gene Manipulation (유전자 재조합 식품의 안전성)

  • 최원상
    • Journal of Food Hygiene and Safety
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    • v.14 no.2
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    • pp.216-225
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    • 1999
  • Safety of present food has been accepted on the basis of extensive use experience for a long time. Many food resources have been developed by traditional techniques without any significant adverse impacts on the safety of food. Recently recombinant DNA techniques are being used to develop new food resources. These techniques enable developers to make specific genetic modifications in food resources that introduce substances that could not be introduced by traditional methods. With these techniques food resources are being to resist pests and disease, to tolerate herbicides, and to have improved characteristics for food preservation and nutritional contents. Because the properties of an organism results from interaction between biochemical pathways controlled by many genes, the genes conferring these traits usually encode directly responsible proteins for the new trait as well as proteins that indirectly modify carbohydrates or lipids in food. Therefore, this kind of food is regarded as new food that has not been existed before, and the safety of the food developed by recombinant DNA techniques should be evaluated upon scientific basis. In this paper, the issues upon safety of the food developed by gene manipulation are diseased in terms of composional changes that can be introduced, potential food safety harzards that might arise, present status of safety regulations in various countries and international organizations, and suggestions for the safety regulation in Korea.

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A Vision Based Bio-Cell Recognition for Biomanipulation with Multiple Views

  • Jang, Min-Soo;Lee, Seok-Joo;Lee, Ho-Dong;Kim, Byung-Kyu;Park, Jong-Oh;Park, Gwi-Tae
    • 제어로봇시스템학회:학술대회논문집
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    • 2003.10a
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    • pp.2435-2440
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    • 2003
  • Manipulation of the nano/micro scale object has been a key technology in biology as the sizes of DNA, chromosome, nucleus, cell and embryo are within such order. For instance, for embryo cell manipulation, the cell injection is performed manually. The operator often spends over a year to carry out a cell manipulation project. Since the typical success rate of such operation is extremely low, automation of such biological cell manipulation has been asked. As the operator spends most of his time in finding the position of cell in the Petri dish and in injecting bio-material to the cell from the best orientation. In this paper, we propose a new strategy and a vision system, by which one can find, recognize and track nucleus, polar body, and zona pellucida of the embryo cell for automatic biomanipulation. The deformable template matching algorithm has been used in recognizing the nucleus and polar body of each cell. Result suggests that it outperforms the conventional methods.

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