• Title/Summary/Keyword: molecular biological techniques

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The expanding reach of the GAL4/UAS system into the behavioral neurobiology of Drosophila

  • Jones, Walton D.
    • BMB Reports
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    • v.42 no.11
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    • pp.705-712
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    • 2009
  • Our understanding of the relationships between genes, brains, and behaviors has changed a lot since the first behavioral mutants were isolated in the fly bottles of the Benzer lab at Caltech (1), but Drosophila is still an excellent model system for studying the neurobiology of behavior. Recent advances provide an unprecedented level of control over fly neural circuits. Efforts are underway to add to existing GAL4-driver lines that permit exogenous expression of genetic tools in small populations of neurons. Combining these driver lines with a variety of inducible UAS lines permits the visualization of neuronal morphology, connectivity, and activity. These driver lines also make it possible to specifically ablate, inhibit, or activate subsets of neurons and assess their roles in the generation of behavioral responses. Here, I will briefly review the extensive arsenal now available to drosophilists for investigating the neuronal control of behavior.

Imaging Gene Expression (유전자 발현 영상기법)

  • Lee, Kyung-Han
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.1
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    • pp.1-9
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    • 2000
  • The rapid progress of molecular genetic methods over the past two decades has necessitated the development of methods to detect and quantify genetic activity within living bodies. Reporter genes provide a rapid and convenient tool to monitor gene expression by yielding a readily measurable phenotype upon expression when introduced into a biological system. Conventional reporter systems, however, are limited in their usefulness for in vivo experiments or human gene therapy because of its invasive nature which requires cell damage before assays can be performed. This offers an unique opportunity for nuclear imaging techniques to develope a novel method for imaging both the location and amount of gene expression noninvasively. Current developments to achieve this goal rely on utilizing either reporter enzymes that accumulate radiolabeled substrates or reporter receptors that bind specific radioligands. This overview includes a brief introduction to the background for such research, a summary of published results, and an outlook for future directions.

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Structure-based Functional Discovery of Proteins: Structural Proteomics

  • Jung, Jin-Won;Lee, Weon-Tae
    • BMB Reports
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    • v.37 no.1
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    • pp.28-34
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    • 2004
  • The discovery of biochemical and cellular functions of unannotated gene products begins with a database search of proteins with structure/sequence homologues based on known genes. Very recently, a number of frontier groups in structural biology proposed a new paradigm to predict biological functions of an unknown protein on the basis of its three-dimensional structure on a genomic scale. Structural proteomics (genomics), a research area for structure-based functional discovery, aims to complete the protein-folding universe of all gene products in a cell. It would lead us to a complete understanding of a living organism from protein structure. Two major complementary experimental techniques, X-ray crystallography and NMR spectroscopy, combined with recently developed high throughput methods have played a central role in structural proteomics research; however, an integration of these methodologies together with comparative modeling and electron microscopy would speed up the goal for completing a full dictionary of protein folding space in the near future.

Pseudoreceptor: Concept and an Overview

  • Kothandan, Gugan;Madhavan, Thirumurthy;Gadhe, Changdev G.;Cho, Seung Joo
    • Journal of Integrative Natural Science
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    • v.3 no.3
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    • pp.162-167
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    • 2010
  • A pseudoreceptor combines structure-based and ligand-based techniques to represent a unifying concept for both receptor mapping and ligand matching. In this molecular modeling approach, there are opportunities to construct the pseudoreceptor models using a set of small molecules. To build a reliable pseudoreceptor model, we need a set of ligand molecules with known affinity (biological activity) to generate 3D bioactive conformation for each of these ligand molecules. Several software packages are available to generate a pseudoreceptor model and this can provide an entry point for structure based drug discovery in cases where receptor structure information is not available. In this review, we presented the concept of pseudoreceptor, as well as discussed about various software packages available to generate a pseudoreceptor model.

Comparison of IHNV Detection Limits by IMS-RT-PCR, Western Blot and ELISA

  • Kim Soo-Jin;Lee Eun-Young;Oh Myung-Joo;Choi Tae-Jin
    • Fisheries and Aquatic Sciences
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    • v.4 no.1
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    • pp.32-38
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    • 2001
  • Several molecular biological techniques have been used to detect virus rapidly and accurately, but these methods have limitations in the early stage of viral infection with very low concentration of virus. We compared the detection limits of IMS-PCR, Western blot and ELISA with infectious hematopoietic necrosis virus OHNV). Four antibodies, rabbit anti-IHNV polyclonal antibody, anti-IHNV nucleocapsid protein monoclonal antibody, anti-IHNV nucleocapsid protein polyclonal antibody, and anti-IHNV glycoprotein polyclonal antibody, were tested to find out the most effective antibody for each method. The detection limit with IMS- PCR was $2\times10^6$ pfu when the viral RNA was extracted before RT-PCR. In the western blot with rabbit anti­IHNV polyclonal antibody one pfu of virus could be detected. In ELISA, 10 pfu of virus particles were detected with the same antibody.

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Electrical Imaging of Thin Film Surface by Scanning Maxwell-stress Microscopy (주사형 맥스웰응력 현미경에 의한 표면의 전기적 이미지)

  • Shin, Hoon-Kyu;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 1998.07d
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    • pp.1508-1510
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    • 1998
  • Recent development of scanning probe microscope techniques has made it possible to investigate, not only microscopic surface topography, but also physical and chemical properties on the nanometer-scale. The scanning Maxwell-stress microscopy (SMM) is surface characterization tool capable of mapping both the surface topography and electrical properties, such as surface potential, surface charge dielectric constant of thin films with a nanometer-scale resolution by means of the AC voltage driven oscillation of metal coated cantilever. In this study, we observed the surface potential distribution and molecular ordering in thin films. We have demonstrated that the SMM can be used for imaging surface potential distribution over the film surface and also be used for detecting surface changes in thin films. This is first step towards the understanding of electrical phenomena in organic and inorganic materials, biological system with SMM.

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Mechanical/Biochemical Analysis of Cell Adhesion Strengthening (세포흡착 거동의 기계적/생화학적 분석)

  • Shin, Heung-Soo
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1455-1457
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    • 2008
  • Cell adhesion is a coordinated process involving initial binding of integrin receptors to extracellular matrix (ECM), recruitment of adhesion proteins, and focal adhesion assembly. The formation of mechanically stable focal adhesion assembly of cells within surrounding ECM is a key parameter to direct numerous cellular functions including cell migration, differentiation, and apotosis. With current cell adhesion assays, it is difficult to understand contributions of each coordinated event on evolution of cell adhesion strengthening since cells spontaneously spread upon their adhesion to the substrate, thus remodeling their cytoskeletal structure. In this presentation, novel approaches for analysis of cell adhesion strengthening process based on the combination of mechanical device, micro-patterned substrates, and molecular biological techniques will be discussed.

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Hyperpolarization: Sensitivity Boost in Magnetic Resonance Spectroscopy and Imaging

  • Ko, Hyeji;Gong, Gyeonghyeon;Jeong, Gijin;Choi, Ikjang;Seo, Hyeonglim;Lee, Youngbok
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.3
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    • pp.124-131
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    • 2015
  • Hyperpolarization methods are the most emerging techniques in the field of magnetic resonance (MR) researches since they make a contribution to overcoming sensitivity limitation of MR spectroscopy and imaging, leading to new fields of researches, real-time in vivo metabolic/molecular imaging and MR analysis of chemical/biological reactions in non-equilibrium conditions. Make use of enormous signal enrichments, it becomes feasible to investigate various chemical and biochemical systems with low ${\gamma}$ nuclei in real-time. This review deals with the theoretical principals of common hyperpolarization methods and their experimental features. In addition, more detailed theories, mechanisms, and applications of dissolution dynamic nuclear polarization (D-DNP) are discussed.

Normalization of Microarray Data: Single-labeled and Dual-labeled Arrays

  • Do, Jin Hwan;Choi, Dong-Kug
    • Molecules and Cells
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    • v.22 no.3
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    • pp.254-261
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    • 2006
  • DNA microarray is a powerful tool for high-throughput analysis of biological systems. Various computational tools have been created to facilitate the analysis of the large volume of data produced in DNA microarray experiments. Normalization is a critical step for obtaining data that are reliable and usable for subsequent analysis such as identification of differentially expressed genes and clustering. A variety of normalization methods have been proposed over the past few years, but no methods are still perfect. Various assumptions are often taken in the process of normalization. Therefore, the knowledge of underlying assumption and principle of normalization would be helpful for the correct analysis of microarray data. We present a review of normalization techniques from single-labeled platforms such as the Affymetrix GeneChip array to dual-labeled platforms like spotted array focusing on their principles and assumptions.

Neurons-on-a-Chip: In Vitro NeuroTools

  • Hong, Nari;Nam, Yoonkey
    • Molecules and Cells
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    • v.45 no.2
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    • pp.76-83
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    • 2022
  • Neurons-on-a-Chip technology has been developed to provide diverse in vitro neuro-tools to study neuritogenesis, synaptogensis, axon guidance, and network dynamics. The two core enabling technologies are soft-lithography and microelectrode array technology. Soft lithography technology made it possible to fabricate microstamps and microfluidic channel devices with a simple replica molding method in a biological laboratory and innovatively reduced the turn-around time from assay design to chip fabrication, facilitating various experimental designs. To control nerve cell behaviors at the single cell level via chemical cues, surface biofunctionalization methods and micropatterning techniques were developed. Microelectrode chip technology, which provides a functional readout by measuring the electrophysiological signals from individual neurons, has become a popular platform to investigate neural information processing in networks. Due to these key advances, it is possible to study the relationship between the network structure and functions, and they have opened a new era of neurobiology and will become standard tools in the near future.