• 제목/요약/키워드: molecular biological techniques

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Recent advances in microfluidic technologies for biochemistry and molecular biology

  • Cho, Soong-Won;Kang, Dong-Ku;Choo, Jae-Bum;Demllo, Andrew J.;Chang, Soo-Ik
    • BMB Reports
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    • 제44권11호
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    • pp.705-712
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    • 2011
  • Advances in the fields of proteomics and genomics have necessitated the development of high-throughput screening methods (HTS) for the systematic transformation of large amounts of biological/chemical data into an organized database of knowledge. Microfluidic systems are ideally suited for high-throughput biochemical experimentation since they offer high analytical throughput, consume minute quantities of expensive biological reagents, exhibit superior sensitivity and functionality compared to traditional micro-array techniques and can be integrated within complex experimental work flows. A range of basic biochemical and molecular biological operations have been transferred to chip-based microfluidic formats over the last decade, including gene sequencing, emulsion PCR, immunoassays, electrophoresis, cell-based assays, expression cloning and macromolecule blotting. In this review, we highlight some of the recent advances in the application of microfluidics to biochemistry and molecular biology.

Comparative Analysis of Gene Expression in the Female Reproductive Organs

  • Kim, Min-Goo;Seo, Hee-Won;Choi, Yo-Han;Lee, Chang-Kyu;Ka, Hak-Hyun
    • 한국수정란이식학회지
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    • 제24권2호
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    • pp.77-87
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    • 2009
  • To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

Unveiling the Diversity of Hydnum in the Republic of Korea with One New Species, Hydnum paucispinum

  • Ji Seon Kim;Wonjun Lee;Changmu Kim;Hanna Park;Chang Sun Kim;Young Woon Lim
    • Mycobiology
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    • 제51권5호
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    • pp.300-312
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    • 2023
  • Hydnum is a genus of ectomycorrhizal fungi belonging to the Hydnaceae family. It is widely distributed across different regions of the world, including North America, Europe, and Asia; however, some of them showed disjunct distributions. In recent years, with the integration of molecular techniques, the taxonomy and classification of Hydnum have undergone several revisions and advancements. However, these changes have not yet been applied in the Republic of Korea. In this study, we conducted an integrated analysis combining the morphological and molecular analyses of 30 specimens collected over a period of approximately 10 years in the Republic of Korea. For molecular analysis, the sequence data of the internal transcribed spacer (ITS) region, the large subunit of nuclear ribosomal RNA gene (nrLSU), and a portion of translation elongation factor 1-a (TEF1) were employed as molecular markers. Through this study, we identified eight species that had previously not been reported to occur in the Republic of Korea, including one new species, Hydnum paucispinum. A taxonomic key and detailed descriptions of the eight Hydnum species are provided in this study.

Visualization of chromatin higher-order structures and dynamics in live cells

  • Park, Tae Lim;Lee, YigJi;Cho, Won-Ki
    • BMB Reports
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    • 제54권10호
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    • pp.489-496
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    • 2021
  • Chromatin has highly organized structures in the nucleus, and these higher-order structures are proposed to regulate gene activities and cellular processes. Sequencing-based techniques, such as Hi-C, and fluorescent in situ hybridization (FISH) have revealed a spatial segregation of active and inactive compartments of chromatin, as well as the non-random positioning of chromosomes in the nucleus, respectively. However, regardless of their efficiency in capturing target genomic sites, these techniques are limited to fixed cells. Since chromatin has dynamic structures, live cell imaging techniques are highlighted for their ability to detect conformational changes in chromatin at a specific time point, or to track various arrangements of chromatin through long-term imaging. Given that the imaging approaches to study live cells are dramatically advanced, we recapitulate methods that are widely used to visualize the dynamics of higher-order chromatin structures.

Glyco-engineering of Biotherapeutic Proteins in Plants

  • Ko, Kisung;Ahn, Mi-Hyun;Song, Mira;Choo, Young-Kug;Kim, Hyun Soon;Ko, Kinarm;Joung, Hyouk
    • Molecules and Cells
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    • 제25권4호
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    • pp.494-503
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    • 2008
  • Many therapeutic glycoproteins have been successfully generated in plants. Plants have advantages regarding practical and economic concerns, and safety of protein production over other existing systems. However, plants are not ideal expression systems for the production of biopharmaceutical proteins, due to the fact that they are incapable of the authentic human N-glycosylation process. The majority of therapeutic proteins are glycoproteins which harbor N-glycans, which are often essential for their stability, folding, and biological activity. Thus, several glyco-engineering strategies have emerged for the tailor-making of N-glycosylation in plants, including glycoprotein subcellular targeting, the inhibition of plant specific glycosyltranferases, or the addition of human specific glycosyltransferases. This article focuses on plant N-glycosylation structure, glycosylation variation in plant cell, plant expression system of glycoproteins, and impact of glycosylation on immunological function. Furthermore, plant glyco-engineering techniques currently being developed to overcome the limitations of plant expression systems in the production of therapeutic glycoproteins will be discussed in this review.

생체영상과 미세가공을 이용한 면역 시스템 연구 (Studying immune system using imaging and microfabrication)

  • 도준상
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2008년도 추계학술대회A
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    • pp.1446-1449
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    • 2008
  • Immune system is composed of multiple cells with distinct functions, and immune responses are orchestrated by complex and dynamic cell-cell interactions. Therefore, each cell behavior and function should be understood under right spatio-temporal context. Studying such complexity and dynamics has been challenging with conventional biological tools. Recent development of new technologies such as state of art imaging instruments and microfabrication techniques compatible with biological systems have provided many exciting opportunities to dissect complex and dynamic immune cell interactions; new microscopy techniques enable us to observe stunning dynamics of immune system in real time. Microfabrication permits us to manipulate microenvironments governing molecular/cellular dynamics of immune cells to study detailed mechanisms of phenomena observed by microscopy. Also, microfabrication can be used to engineer microenvironments optimal for specific imaging techniques. In this presentation, I am going to present an example of how these two techniques can be combined to tackle challenging problems in immunology. Obviously, this strategy can readily be applied to many different fields of biology other than immunology.

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Neural and Genetic Basis of Evasion, Approach and Predation

  • Park, Seahyung;Ryoo, Jia;Kim, Daesoo
    • Molecules and Cells
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    • 제45권2호
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    • pp.93-97
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    • 2022
  • Evasion, approach and predation are examples of innate behaviour that are fundamental for the survival of animals. Uniting these behaviours is the assessment of threat, which is required to select between these options. Far from being comprehensive, we give a broad review over recent studies utilising optic techniques that have identified neural circuits and genetic identities underlying these behaviours.

Looking Inside the Cell for Mechanisms of Immunotoxicity: Experimental Design and Approaches Aimed Toward Elucidation of 2,3,7,8-Tetrachlor- dibenzo-p-dioxin-mediated B Cell Dysfunction

  • Norbert E. Kaminski;Kang, Jong-Soon
    • Toxicological Research
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    • 제17권
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    • pp.205-210
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    • 2001
  • One of the major focuses and perhaps the greatest challenges during the past decade in the discipline of immunotoxicology has been the elucidation of the molecular mechanisms responsible for immunotoxicity by specific agents. Much is currently understood about the basic underlying intracellular processes that control leukocyte effector function. This fundamental information in cell biology can now be applied toward developing systematic approaches, through the application of cell and molecular biology techniques, to identify the intracellular targets and processes disrupted by immunotoxicants. The objective of this paper is two fold. First to discuss fundamental principles of experimental design aimed at elucidation of cellular mechanisms in immunotoxicology; and second to discuss the application of molecular biology techniques in characterizing the mechanism of TCDD-induced B cell dysfunction as a working example.

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Synergistic Ensemble of Optogenetic Actuators and Dynamic Indicators in Cell Biology

  • Kim, Jihoon;Heo, Won Do
    • Molecules and Cells
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    • 제41권9호
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    • pp.809-817
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    • 2018
  • Discovery of the naturally evolved fluorescent proteins and their genetically engineered biosensors have enormously contributed to current bio-imaging techniques. These reporters to trace dynamic changes of intracellular protein activities have continuously transformed according to the various demands in biological studies. Along with that, light-inducible optogenetic technologies have offered scientists to perturb, control and analyze the function of intracellular machineries in spatiotemporal manner. In this review, we present an overview of the molecular strategies that have been exploited for producing genetically encoded protein reporters and various optogenetic modules. Finally, in particular, we discuss the current efforts for combined use of these reporters and optogenetic modules as a powerful tactic for the control and imaging of signaling events in cells and tissues.