• Title/Summary/Keyword: differential display analysis

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Analysis of Gene Expression in Benzo[a]pyrene-exposed Sebastes schlegeli using Differential Display Polymerase Chain Reaction (DD-PCR을 이용한 벤조피렌 노출 조피볼락의 차등 발현 유전자 분석)

  • Yum Seungshic;Woo Seonock;Choi Eunseok;Kim Sojung;Oh Rora;Lee Sukchan;Lee Taek Kyun
    • Environmental Analysis Health and Toxicology
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    • v.20 no.1
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    • pp.67-73
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    • 2005
  • 오염물질의 노출에 의해 발현이 변화되는 유전자의 발굴은 외부환경 자극에 대한 적응이나 반응의 메커니즘을 알아내는뎨 중요한 정보를 제공하며, 오염물질에 반응하는 유전자는 환경오염을 감지하는 분자 마커로 개발될 수 있다. DD-PCR 기법은 차등 발현 유전자들을 발굴해내기 위한 유용한 방법으로 사용되어 왔고, 본 연구는 이 방법을 이용하여 벤조피렌에 반응하는 조피볼락 유전자들의 발굴을 목적으로 진행되었다. 간조직에서 추출한 RNA로부터 벤조피렌의 노출에 의해 발현 양이 달라진 12개의 클론을 발굴하였고, 그 염기서 열을 분석하였다. 또한 벤조피렌의 노출시간을 각각 6, 12, 24시간으로 달리한 조피볼락에서 12개의 클론 중 4개의 클론에 대해 northern blot 분석이 실시되었으며, 이들 모두 노출시간에 따 라 발현양이 증가 또는 감소하는 것이 확인되었다. 본 연구결과는 오염물질의 영향에 의한 유전자들의 발현에 관한 전반적인 지식을 제공하였고, 나아가 환경오염이나 외부 스트레스를 감지해 낼 수 있는 바이오마커의 개발을 위한 첫 단계로서의 정보를 제공할 수 있을 것으로 생각된다.

Effect of CaO on the antibacterial property of zinc borosilicate glasses (Zinc borosilicate 유리의 CaO 첨가에 따른 항균력 개선 효과 검증)

  • Minsung Hwang;Jaeyeop Chung
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.33 no.5
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    • pp.187-190
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    • 2023
  • In this study, antibacterial glasses were developed by the addition of CaO in ZnO-Na2O-B2O3-SiO2 glass system. The effect of the addition of CaO on the thermal properties, dissolution properties, surface zeta potential and antibacterial activity were analyzed. Differential thermal analysis (DTA) analysis was performed to analyze the thermophysical properties of 30ZnO-xCaO-20Na2O-30B2O3-(20-x)SiO2 (x = 0, 2, 4, 6, 8, 10 mol%). It was confirmed that the glass transition temperature decreased as the CaO content increased. The amount of released Zn2+ ions and surface zeta potential of glass samples increased with increasing CaO concentration. For these reasons, the antibacterial activity was dramatically improved. By the addition of CaO, we could successfully develop an antibacterial glass with 99.9 % antibacterial activity against both Escherichia coli and Staphylococcus aureus.

Toward Functional Genomics of Plant-Pathogen Interactions: Isolation and Analysis of Defense-related Genes of Rot Pepper Expressed During Resistance Against Pathogen

  • Park, Do-Il;Lee, Sang-Hyeob
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.63-67
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    • 2002
  • To understand plant-pathogen interactions, a complete set of hot pepper genes differentially expressed against pathogen attack was isolated. As an initial step, hundreds of differentially expressed cDNAS were isolated from hot pepper leaves showing non-host resistance against bacterial plant pathogens (Xanthomonas campestris pv. glycines and Pseudomonas syringae pv. syringae) using differential display reverse transcription polymerase chain reaction (DDDRT-PCR) technique. Reverse Northern and Northern blot analyses revealed that 50% of those genes were differentially expressed in pepper loaves during non-host resistance response. Among them, independent genes without redundancy were micro-arrayed for further analysis. Random EST sequence database were also generated from various CDNA libraries including pepper tissue specific libraries and leaves showing non-host hypersensitive response against X. campestris pv. glycines. As a primary stage, thousands of cDNA clones were sequenced and EST data were analyzed. These clones are being spotted on glass slide to study the expression profiling. Results of this study may further broaden knowledge on plant-pathogen interactions.

A Trial of Screening of Genes Involved in Odontoblasts Differentiation from Human Dental Pulp Stem Cells

  • Park, Yoon-Kyu;Kim, Hyun-Jin
    • International Journal of Oral Biology
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    • v.37 no.4
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    • pp.167-173
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    • 2012
  • This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

Development of AVL-GIS System Using IDGPS and Wireless Communication Techniques (IDGPS 와 무선통신을 이용한 AVL-GIS 시스템개발)

  • 안충현;양종윤;최종현
    • Spatial Information Research
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    • v.7 no.2
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    • pp.209-221
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    • 1999
  • In this research, AVL-GIS(Automatic Vehicle Location System linked with Geographic Information System) system was developed using integration of core techniques of GIS engine written by Java language, GOS(Global Positioning System) and wireless telecommunication interfacing techniques. IDGPS(Inverted differential GPS) techniques was employed to estimate accurate position of mobile vehicle and to supervise their path from AVL-GLS control center system. Between mobile vehicle and AVL-GLS control center system which has spatial data analysis function, road network and rleate ddata base were connected wireless phone to communicate for position an dmessage in real time. The developed system from this research has more enhanced GIS functions rather than previous AVL oriented system which has MDT for message display and voice communication only. This system can support build-up application system such as fleet management like bus, taxi, truck, disaster and emergency and monitoring of transportation status for customer s order via web browser in filed of EC/CALS in low cost.

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Second Order Nonlinear Optical Polyimides Containing Organic Chromophores with an Oxadiazole Segment (옥사디아졸 결합의 유기 발색단이 도입된 이차비선형 광학 이미드 고분자)

  • Do, Jung Yun;Kim, Bong Gun;Kwon, Ji-Yun
    • Applied Chemistry for Engineering
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    • v.18 no.1
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    • pp.77-83
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    • 2007
  • It is essential that second order nonlinear optical materials have low optical propagation losses in the wavelengths of second harmonic generation for practical applications in waveguides. Three dipolar chromophores substituted with nitro, cyano, and alkyl sulfone as an electron withdrawing group were prepared. The UV-Vis absorption spectra of the cyano and alkylsulfone chromophores showed a blue-shift compared to the nitro chromophore. The introduction of oxadiazole segment in the chromophore structure led to similar spectral shift. The blue-shift can produce low optical loses at second harmonics. The chromophores were successfully attached to a polyimide, yielding side chain polymers. The nonlinear optical property of the prepared optical polymers was determined by measuring electro-optic coefficient at 1.55 mm. The polymers exhibited high glass transition temperature of over $185^{\circ}C$ and thermal stability to $300^{\circ}C$ through differential scanning calorimeter analysis and thermal gravimetric analysis.

Identification of novel susceptibility genes associated with bone density and osteoporosis in Korean women

  • Bo-Young Kim;Do-Wan Kim;Eunkuk Park;Jeonghyun Kim;Chang-Gun Lee;Hyun-Seok Jin;Seon-Yong Jeong
    • Journal of Genetic Medicine
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    • v.19 no.2
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    • pp.63-75
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    • 2022
  • Purpose: Osteoporosis is a common calcium and metabolic skeletal disease which is characterized by decreased bone mass, microarchitectural deterioration of bone tissue and impaired bone strength, thereby leading to enhanced risk of bone fragility. In this study, we aimed to identify novel genes for susceptibility to osteoporosis and/or bone density. Materials and Methods: To identify differentially expressed genes (DEGs) between control and osteoporosis-induced cells, annealing control primer-based differential display reverse-transcription polymerase chain reaction (RT-PCR) was carried out in pre-osteoblast MC3T3-E1 cells. Expression levels of the identified DEGs were evaluated by quantitative RT-PCR. Association studies for the quantitative bone density analysis and osteoporosis case-control analysis of single nucleotide polymorphism (SNPs) were performed in Korean women (3,570 subjects) from the Korean Association REsource (KARE) study cohort. Results: Comparison analysis of expression levels of the identified DEGs by quantitative RT-PCR found seven genes, Anxa6, Col5a1, Col6a2, Eno1, Myof, Nfib, and Scara5, that showed significantly different expression between the dexamethason-treated and untreated MC3T3-E1 cells and between the ovariectomized osteoporosis-induced mice and sham mice. Association studies revealed that there was a significant association between the SNPs in the five genes, ANXA6, COL5A1, ENO1, MYOF, and SCARA5, and bone density and/or osteoporosis. Conclusion: Using a whole-genome comparative expression analysis, gene expression evaluation analysis, and association analysis, we found five genes that were significantly associated with bone density and/or osteoporosis. Notably, the association P-values of the SNPs in the ANXA6 and COL5A1 genes were below the Bonferroni-corrected significance level.

Characterization of Pathogenesis and Plant Defence-related Genes Against Potato virus X infection empolying Potato X virus expresssin vector

  • Park, Mi-Ri;Kwon, Sun-Jung;Kim, Kook-Hyung
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.74.1-74
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    • 2003
  • Differential display (DD) of mRNA is a technique in which mRNA species expressed by a cell population are reverse transcribed and then amplified by many separate polymerase chain reactions (PCR). Using DD-RT-PCR we obtained many genes that expressed differentially in healthy and PVX-infected Nicotiana benthamima, using total RNAs extracted from healthy and PVX-infected N. benthamiana plants. Three hundred and twenty-five DNA fragments isolated from DD-RT-PCR were cloned and sequenced for further characterization. Several host genes including SKPI-like protein, heat shock transcription factor and Avr9/Cf-9 rapidly elicited protein were selected to obtain full-length open reading frame and to characterize their potential involvement in virus disease development and/or host's defense against virus infection employing PVX-based expression vector. Transcrips from wild-type and clones containing each selected gene were inoculated onto N. benthamiana Levels of virus replication were confirmedby RT-PCR and RNA blot analysis, Expression profiles and potential role(s) of selected genes upon PVX infection will be discussed.

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A Fluorescence-based cDNA-AFLP Method for Identification of Differentially Expressed Genes

  • Park, Sook-Young;Jwa, Nam-Soo;Chi, Myoung-Hwan;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.184-188
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    • 2009
  • Identification of differently expressed genes under specific tissues and/or environments provides insights into the nature and underlying mechanisms of cellular processes. Although cDNA-AFLP (Amplified Fragment Length Polymorphism) is a powerful method for analyzing differentially expressed genes, its use has been limited to the requirement of radioactive isotope use and the difficulty of isolating the bands of interest from a gel. Here, we describe a modified method for cDNA-AFLP that uses a fluorescence dye for detection and isolation of bands directly from a small size polyacrylamide gel. This method involves three steps: (i) preparation of cDNA templates, (ii) PCR amplification and differential display, and (iii) identification of differentially expressed genes. To demonstrate its utility and efficiency, differentially expressed genes during vegetative growth and appressorial development of Magnaporthe oryzae were analyzed. This method could be applied to compare gene expression profiles in a diverse array of organisms.

CLONGING OF GENES EXPRESSED UPON FLORAL INDUCTION IN PHARBITIS COTYLEDONS

  • Kim, Kang-Chang;Hur, Yoon-Kang;Maeng, Jue-Son
    • Journal of Photoscience
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    • v.5 no.3
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    • pp.131-135
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    • 1998
  • Using differential display reverse transcription technique, the present study attempted to isolate and characterize genes specifically expressed in cotyledons of Pharbitis nil Choisy cv. Violet during floral induction. A total of 107 bands specific to the inductive condition were initially obtained with 80 primer sets of 20 different arbitrary primers combined with 4 kinds of T12MN. In northern blot analysis with reamplified cDNAs as probes, three cDNAs were detected to be expressed specificcally in the induced cotyledon tissues, and designated PnFL-1, PnFL-2 and PnFL-3 , the size of which were 228 bp, 317 bp and 272 bp, respectively. A search for sequences similar to the decuced amino acid sequences was conducted using GenBank and EMBL database ; seequence encoded by PnFL-1 had 29% identity with the clone of Arabidopsis thaliana similiar to reverse trascriptase (Genbank Acc. N0.3047086), PnFL-2 shared 50% identity with hydroxiyproline-rich glycoprotein of Glycine max(GenBank Acc. No.347455), and PnFL-3 had 46% identity with TAMU 4. Thaliana genomic clone T23E16 (GenBank Acc. No.B67574). None of them was known gene in the plant system up to date, implying that the fragments may comprise parts of genes which are associated with the floral induction in Pharbitis nil.

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