• 제목/요약/키워드: Differential display analysis

검색결과 102건 처리시간 0.023초

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

  • 염승식;우선옥;최은석;김소정;오로라;이석찬;이택견
    • Environmental Analysis Health and Toxicology
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    • 제20권1호
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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.

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

  • 황민성;정재엽
    • 한국결정성장학회지
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    • 제33권5호
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    • pp.187-190
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    • 2023
  • 본 연구에서는 ZnO-Na2O-B2O3-SiO2 계 유리에 CaO의 첨가에 따른 유리의 열적, 화학적 특성, 표면 제타전위 및 항균특성을 분석하였다. 유리 조성에 따른 열적 특성은 DTA 분석을 통해 확인하였고, 30ZnO-xCaO-20Na2O-30B2O3-(20-x)SiO2 (x = 0, 2, 4, 6, 8, 10 mol%)계 유리에서 CaO 함량이 증가함에 따라 유리전이온도가 줄어드는 것을 확인하였다. CaO 함량이 늘어날수록 유리 구조가 약화됨에 따라 더 많은 Zn2+ 이온이 용출되었고, 알칼리 및 알칼리 토류의 초기 급속한 용출로 인해 유리의 표면 제타전위가 증가함을 확인하였다. 이러한 이유로 유리의 항균활성 또한 급격하게 개선됨을 확인하였으며, 대장균(gram negative)과 황색포도상구균(gram positive) 모두에 대해 99.9 % 항균 활성을 갖는 항균 유리를 개발할 수 있었다.

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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    • 제18권2호
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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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    • 제37권4호
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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.

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

  • 안충현;양종윤;최종현
    • Spatial Information Research
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    • 제7권2호
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    • pp.209-221
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    • 1999
  • 자바언어로 구현된 GIS 공간정보분서엔진, GPS, 휴대전화를 이용한 무선 데이터 통신을 연계하여 AVL-GIS 시스템을 개발하였으며, 프로토 타입으로서 긴급 상황 관제 센터로 가정한 통합안전관리시스템(경찰, 119 , 소방서)제 적용, 구현하였다. 이동차량이 정확한 위치의 추적을 위하여 IDGPS(Inverted Differential GPS) 방법을 사용하였으며, 휴대전화를 이용하여 GIS 분석기능과 도로 데이터 및 이와 관련된 공간정보를 갖고 있는 중앙 관제시스템을 실시간으로 연결하여 위치정보 및 메시지를 송수신할 수 있도록 하였다. 본 연구에서 개발된 시스템은 기존의 AVL용 시스템보다 GIS 분석기능이 강화되어 있으며, 비교적 저렴한 가격으로 물류정보, 재난 및 안전관리, 버스나 택시와 같은 차량관리와 전자상거래에서 인터넷을 통한 화물의 위치 확인등과 같은 응용시스템을 구축할 수 있다.

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

  • 도정윤;김봉건;권지윤
    • 공업화학
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    • 제18권1호
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    • pp.77-83
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    • 2007
  • 2차 비선형 광학 소재는 광 도파로 응용을 위해 조화파 파장영역에서 낮은 광 진행 손실을 가져야 한다. 이를 위하여 분자에 전자 당김 작용기로 니트로기, 시안기 및 알킬슬폰기가 각각 도입된 세 가지의 쌍극자형 발색단 물질이 합성되었다. 시안기 및 슬폰기를 갖는 발색단의 자외-가시 흡수 스펙트럼은 니트로기를 갖는 발색단에 비해 단파장으로 이동하였다. 또한, 크로모포 분자에 옥사디아졸 연결기를 도입한 결과 광흡수 스펙트럼이 단파장으로 이동하는 유사한 특성을 관찰하였다. 이러한 단파장으로 이동하는 특성은 2차 조화파의 낮은 광손실을 유도할 것이다. 합성된 크로모포는 이미드 고분자에 곁사슬기 형태로 도입하였다. 합성된 고분자의 비선형 광학 성질은 $1.55{\mu}m$ 파장 영역에서 전기광학계수를 측정하고 변환을 통하여 결정하였다. 시차열량 분석계와 열중량 분석계를 이용하여 이들 고분자의 물성 측정을 진행한 결과 $185^{\circ}C$ 이상의 높은 유리전이 온도와 $300^{\circ}C$까지 열적으로 안정함을 보였다.

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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    • 제19권2호
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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
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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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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    • 제25권2호
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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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    • 제5권3호
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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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