• Title/Summary/Keyword: Molecular approach

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Preparation of Nanostructures Using Layer-by-Layer Assembly and Applications (층상자기조립법을 이용한 나노구조체의 제조와 응용)

  • Cho, Jin-Han
    • Journal of the Korean Vacuum Society
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    • v.19 no.2
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    • pp.81-90
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    • 2010
  • We introduce a novel and versatile approach for preparing self-assembled nanoporous multilayered films with antireflective properties. Protonated polystyrene-block-poly (4-vinylpyrine) (PS-b-P4VP) and anionic polystyrene-block-poly (acrylic acid) (PS-b-PAA) block copolymer micelles (BCM) were used as building blocks for the layer-by-layer assembly of BCM multilayer films. BCM film growth is governed by electrostatic and hydrogen-bonding interactions between the oppositely BCMs. Both film porosity and film thickness are dependent upon the charge density of the micelles, with the porosity of the film controlled by the solution pH and the molecular weight (Mw) of the constituents. PS7K-b-P4VP28K/PS2K-b-PAA8K films prepared at pH 4 (for PS7K-b-P4VP28K) and pH 6 (for PS2K-b-PAA8K) are highly nanoporous and antireflective. In contrast, PS7K-b-P4VP28K/PS2K-b-PAA8K films assembled at pH 4/4 show a relatively dense surface morphology due to the decreased charge density of PS2K-b-PAA8K. Films formed from BCMs with increased PS block and decreased hydrophilic block (P4VP or PAA) size (e.g., PS36K-b-P4VP12K/PS16K-b-PAA4K at pH 4/4) were also nanoporous. Furthermore, we demonstrate that the nanostructured electrochemical sensors based on patterning methods show the electrochemical activities. Anionic poly(styrene sulfonate) (PSS) layers were selectively and uniformly deposited onto the catalase (CAT)-coated surface using the micro-contact printing method. The pH-induced charge reversal of catalase can provide the selective deposition of consecutive PE multilayers onto patterned PSS layers by causing the electrostatic repulsion between next PE layer and catalase. Based on this patterning method, the hybrid patterned multilayers composed of platinum nanoparticles (PtNP) and catalase were prepared and then their electrochemical properties were investigated from sensing $H_2O_2$ and NO gas. This study was based on the papers reported by our group. (J. Am. Chem. Soc. 128, 9935 (2006); Adv. Mater. 19, 4364 (2007); Electro. Mater. Lett. 3, 163 (2007)).

Establishing Effective Screening Methodology for Novel Herbicide Substances from Metagenome (신규 제초활성 물질 발굴을 위한 메타게놈 스크리닝 방법 연구)

  • Lee, Boyoung;Choi, Ji Eun;Kim, Young Sook;Song, Jae Kwang;Ko, Young Kwan;Choi, Jung Sup
    • Weed & Turfgrass Science
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    • v.4 no.2
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    • pp.118-123
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    • 2015
  • Metagenomics is a powerful tool to isolate novel biocatalyst and biomolecules directly from the environmental DNA libraries. Since the metagenomics approach bypasses cultivation of microorganisms, un-cultured microorganisms that are majority of exists can be the richest reservoir for natural products discovery. To discover novel herbicidal substances from soil metagenome, we established three easy, simple and effective high throughput screening methods such as cucumber cotyledon leaf disc assay, microalgae assay and seed germination assay. Employing the methods, we isolated two active single clones (9-G1 and 9-G12) expressing herbicidal activity which whitened leaf discs, inhibited growth of microalgae and inhibited root growth of germinated Arabidopsis seeds. Spraying butanol fraction of the isolated active clones' culture broth led to growth retardation or desiccation of Digitalia sanguinalis (L) Scop. in vivo. These results represent that the screening methods established in this study are useful to screen herbicidal substances from metagenome libraries. Further identifying molecular structure of the herbicidal active substances and analyzing gene clusters encoding synthesis systems for the active substances are in progress.

Surveillance of Acanthamoeba spp. and Naegleria fowleri in environmental water by using the duplex real-time PCR (Duplex real-time PCR을 이용한 수계 중 가시아메바와 파울러자유아메바 조사)

  • Kim, Min-jeong;Lee, Gyu-Cheol;Kim, Kunwoo;Lee, Hyunji;Kim, Min Young;Seo, Dae Keun;Lee, Jeong Yeob;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.98-104
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    • 2018
  • Naegleria fowleri and Acanthamoeba spp. are free-living amoebas that are widely distributed in natural environments. Although uncommon, infection with these protozoans can cause fatal disease in humans and animals. In this study, in order to select the appropriate method to survey Naegleria fowleri and Acanthamoeba spp. in water samples, four molecular biology techniques and one commercially available kit for real-time PCR were compared. The results indicated that the duplex real-time PCR was the most sensitive, and could be used to simultaneously detect two different free-living amoebas. Using the duplex real-time PCR approach, the two free-living amoebas were surveyed in three local streams in Daejeon, Republic of Korea. The concentrated free-living amoebas were inoculated onto non-nutrient agar plates which had been spread with heat-inactivated Escherichia coli and incubated for 5~7 days. After incubation, gDNA was extracted and used as the template for amplification by duplex real-time PCR. Acanthamoeba spp. and N. fowleri was detected from ten (83.3%) and two (16.6%) of the twelve samples, respectively. As these two free-living amoebas can be fatal, continuous surveillance is needed to track their distribution in the aquatic environment for the drinking water safety.

Comparative Analysis of Peptide Nucleic Acid (PNA)-Mediated Real-Time PCR Clamping and DNA Direct Sequencing for EGFR Mutation Detection (EGFR 돌연변이 검출에 있어 PNA-Mediated Real-Time PCR Clamping과 직접 염기서열 분석법의 비교 분석)

  • Kim, Hee-Joung;Kim, Wan-Seop;Shin, Kyeong-Cheol;Lee, Gwan-Ho;Kim, Mi-Jin;Lee, Jeong-Eun;Song, Kyu-Sang;Kim, Sun-Young;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.1
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    • pp.21-27
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    • 2011
  • Background: Although the gold standard method for research trials on epidermal growth factor receptor (EGFR) mutations has been direct sequencing, this approach has the limitations of low sensitivity and of being time-consuming. Peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping is known to be a more sensitive detection tool. The aim of this study was to compare the detection rate of $EGFR$ mutation and EGFR-tyrosine kinase inhibitor (TKI) responsiveness according to $EGFR$ mutation status using both methodologies. Methods: Clinical specimens from 112 NSCLC patients were analyzed for $EGFR$ mutations in exons 18, 19, 20, and 21. All clinical data and tumor specimens were obtained from 3 university hospitals in Korea. After genomic DNA was extracted from paraffin-embedded tissue specimens, both PNA-mediated PCR clamping and direct-sequencing were performed. The results and clinical response to $EGFR$-TKIs were compared. Results: Sequencing revealed a total of 35 (22.9%) mutations: 8 missense mutations in exon 21 and 26 deletion mutations in exon 19. PNA-mediated PCR clamping showed the presence of genomic alterations in 45 (28.3%) samples, including the 32 identified by sequencing plus 13 additional samples (6 in exon 19 and 7 in exon 21). Conclusion: PNA-mediated PCR clamping is simple and rapid, as well as a more sensitive method for screening of genomic alterations in $EGFR$ gene compared to direct sequencing. This data suggests that PNA-mediated PCR clamping should be implemented as a useful screening tool for detection of $EGFR$ mutations in clinical setting.

Studies on the Optical and the Electrical Characterization of Organic Electroluminescence Devices of Europium Complex Fabricated with PVD(Physical Vopor Deposition) Technique (진공 증착법에 의하여 제작한 Europium complex 유기 박막 전기발광소자의 광학적.전기적 특성에 관한 연구.)

  • Lee, Myeong-Ho;Lee, Han-Seong;Kim, Yeong-Gwan;Kim, Jeong-Su
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.48 no.5
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    • pp.285-295
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    • 1999
  • Electroluminescent(EL) devices based on organic materials have been of great interest due to their possible applications for large-area flat-panel displays. They are attractive because of their capability of multi-color emission, and low operation voltage. An approach to realize such device characteristics is to use active layers of lanthanide complexes with their inherent extremely sharp emission bands in stead of commonly known organic dyes. In general, organic molecular compounds show emission due to their $\pi$-$\pi*$ transitions resulting in luminescence bandwidths of about 80 to 100nm. Spin statistic estimations lead to an internal quantum efficiency of dye-based EL devices limited to 25%. On the contrary, the fluorescence of lanthanide complexes is based on an intramolecular energy transfer from the triplet of the organic ligand to the 4f energy states of the ion. Therefore, theoretical internal quantum efficiency is principally not limited. In this study, Powders of TPD, $Eu(TTA)_3(phen) and AlQ_3$ in a boat were subsequently heated to their sublimation temperatures to obtain the growth rates of 0.2~0.3nm/s. Organic electrolumnescent devices(OELD) with a structure of $glass substrate/ITO/Eu(TTA)_3(phen)/AI, glass substrate/ITO/TPD/Eu(TTA)_3(phen)/AI and glass substrate/ITO/TPD/Eu(TTA)_3(phen)/AIQ_3AI$ structures were fabricated by vacuum evaporation method, where aromatic diamine(TPD) was used as a hole transporting material, $Eu(TTA)_3(phen)$ as an emitting material, and Tris(8-hydroxyquinoline)Aluminum$(AlQ_3)$ as an electron transporting layer. Electroluminescent(EL) and current density-voltage(J-V) characteristics of these OELDs with various thickness of $Eu(TTA)_3(phen)$ layer were investigated. The triple-layer structure devices show the red EL spectrum at the wavelength of 613nm, which is almost the same as the photoluminescent(PL) spectrum of $Eu(TTA)_3(phen)$.It was found from the J-V characteristics of these devices that the current density is not dependent on the applied field, but on the electric field.

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Analysis of Human Skeletal Remains of the Joseon Dynasty from Hwamyeong-dong, Busan: A Molecular Genetic Approach (분자유전학적 접근을 통한 조선시대 사람뼈의 분석 - 부산 화명동 조선시대 분묘군 출토 사람뼈를 중심으로 -)

  • Kim, Sue Hoon;Cho, Eun Min;Kim, Yun-Ji;Choe, Hyeongoo;Kang, Soyeong
    • Journal of Conservation Science
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    • v.34 no.1
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    • pp.1-9
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    • 2018
  • The analysis of ancient DNA extracted from archaeological bones has become an important research tool in palaeogenetics and anthropology. Eight human skeletal remains of the Joseon dynasty, excavated from Hwamyeong-dong, were used in this study. DNA was extracted from bone powder using a silica-based protocol. The isolated DNA was analyzed by the sequencing variation of hyper-variable region of the mitochondrial DNA. In the present study, 3 human remains were identified into mtDNA haplogroups including the A 5a, D4a, and M4"67+16311 groups, using HaploGrep 2 program. The identified haplotypes of the 3 samples have been confirmed that the specimens in the tombs were not related by the maternal line. This is the first analysis of human skeletal remains of the Joseon dynasty excavated in Busan. Date from the analysis of human remains from the Joseon dynasty are considered as the basis for understanding the genetic relationship between modern and ancient humans of the Korean peninsula.

Molecular Genetic Classification of Hypsizigus marmoreus and Development of Strain-specific DNA Markers (느티만가닥버섯의 분자유전학적 분류 및 품종특이적 DNA 마커 탐색)

  • Lim, Yun-Jeong;Lee, Chang-Yun;Park, Jeong-Eun;Kim, Sang-Woo;Lee, Hyun-Sook;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.34-39
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    • 2010
  • We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

Gene Discovery Analysis from Mouse Embryonic Stem Cells Based on Time Course Microarray Data

  • Suh, Young Ju;Cho, Sun A;Shim, Jung Hee;Yook, Yeon Joo;Yoo, Kyung Hyun;Kim, Jung Hee;Park, Eun Young;Noh, Ji Yeun;Lee, Seong Ho;Yang, Moon Hee;Jeong, Hyo Seok;Park, Jong Hoon
    • Molecules and Cells
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    • v.26 no.4
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    • pp.338-343
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    • 2008
  • An embryonic stem cell is a powerful tool for investigation of early development in vitro. The study of embryonic stem cell mediated neuronal differentiation allows for improved understanding of the mechanisms involved in embryonic neuronal development. We investigated expression profile changes using time course cDNA microarray to identify clues for the signaling network of neuronal differentiation. For the short time course microarray data, pattern analysis based on the quadratic regression method is an effective approach for identification and classification of a variety of expressed genes that have biological relevance. We studied the expression patterns, at each of 5 stages, after neuronal induction at the mRNA level of embryonic stem cells using the quadratic regression method for pattern analysis. As a result, a total of 316 genes (3.1%) including 166 (1.7%) informative genes in 8 possible expression patterns were identified by pattern analysis. Among the selected genes associated with neurological system, all three genes showing linearly increasing pattern over time, and one gene showing decreasing pattern over time, were verified by RT-PCR. Therefore, an increase in gene expression over time, in a linear pattern, may be associated with embryonic development. The genes: Tcfap2c, Ttr, Wnt3a, Btg2 and Foxk1 detected by pattern analysis, and verified by RT-PCR simultaneously, may be candidate markers associated with the development of the nervous system. Our study shows that pattern analysis, using the quadratic regression method, is very useful for investigation of time course cDNA microarray data. The pattern analysis used in this study has biological significance for the study of embryonic stem cells.

A Study on Elution Behavior of Polystyrene Copolymers in Gel Permeation Chromatography (겔 투과 크로마토그래피에서 폴리스티렌 혼성중합체들의 용리거동에 관한 연구)

  • Lee Dai Woon;Eum Chul Hun
    • Journal of the Korean Chemical Society
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    • v.36 no.1
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    • pp.87-94
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    • 1992
  • The elution behavior of polystyrenes(PS), polymethylmethacrylates (PMMA), polybutadienes(PB), PS-PMMA(SM) block copolymers and PS-PB star shaped copolymers on the cross-linked polystyrene gels was studied. An interpretation was proposed for the plots of log hydrodynamic volume versus retention volume of solutes in the mobile phases such as tetrahydrofuran, toluene, chloroform, methylene chloride and tetrahydrofuran-cyclohexane mixture. In order to predict the retention of solutes from their physical properties, multiple stepwise regression analysis was applied to obtain the correlation. The distribution coefficients($K_p$) of solute-gel interactions in GPC for homopolymers and PS copolymers were also obtained in terms of network-limited separation mechanism. In the cases of PS and PB, $K_p$ values approach unity, while $K_p$ values for PMMA decrease as MW increase in the good solvent, but in poor solvent, $K_p$ values increase as MW increase. $K_p$ values of PS copolymers are dependent on their MW and composition, therefore, morohology of SM block copolymer is predicted to be random phase. A single universal plot of log[η]M vs. $(V_r-V_o)/K_p$

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Application and Utilization of Environmental DNA Technology for Biodiversity in Water Ecosystems (수생태계 생물다양성 연구를 위한 환경유전자(environmental DNA) 기술의 적용과 활용)

  • Kwak, Ihn-Sil;Park, Young-Seuk;Chang, Kwang-Hyeon
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.151-155
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    • 2021
  • The application of environmental DNA in the domestic ecosystem is also accelerating, but the processing and analysis of the produced data is limited, and doubts are raised about the reliability of the analyzed and produced biological taxa identification data, and the sample medium (target sample, water, air, sediment, Gastric contents, feces, etc.) and quantification and improvement of analysis methods are also needed. Therefore, in order to secure the reliability and accuracy of biodiversity research using the environmental DNA of the domestic ecosystem, it is a process of actively using the database accumulated through ecological taxonomy and undergoing verification procedures, and experts verifying the resolution of the data increased by gene sequence analysis. This is absolutely necessary. Environmental DNA research cannot be solved only by applying molecular biology technology, and interdisciplinary research cooperation such as ecology-taxa identification-genetics-informatics is important to secure the reliability of the produced data, and researchers dealing with various media can approach it together. It is an area in desperate need of an information sharing platform that can do this, and the speed of development will proceed rapidly, and the accumulated data is expected to grow as big data within a few years.