• Title/Summary/Keyword: Differential Display

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Molecular Cloning of Differentially Expressed Genes in First Trap Leaf of Dionaea muscipula by Fluorescent Differential Display (형광 Differential Display법에 의한 파리지옥풀 포충잎트랩 특이발현 유전자 탐색)

  • Kang, Kwon-Kyoo;Lee, Keun-Hyang;Park, Jin-Heui;Hong, Kyong-Ei
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.307-313
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    • 2003
  • Fluorescent differential display (FDD) is a method for identifying differentially expressed genes in eukaryotic cells. The mRNA FDD technology works by systematic amplification of the 3' terminal regions of mRNAs. This method involve the reverse transcription using anchored primers designed to bind 5'boundary of the poly A tails, followed by polymerase chain reaction (PCR) amplification with additional upstream primers of arbitrary sequences. The amplified cDNA subpopulations are separated by denaturing polyacrylamide electrophoresis. To identify the genes involved in the development of first trap leaf, we applied a FDD method using mRNAs from leaf base, first trap leaf and flower tissue, respectively. We screened several genes that expressed specifically in first trap leaf. Nucleotide sequence analysis of these genes revealed that these were protease inhibitor (PI), myo-inositol-1-phosphate synthase and lipocalin-type prostaglandin D synthase. Northern blot analysis showed that these genes were expressed specifically in first trap leaf (in vivo and in vitro). FDD could prove to be useful for simultaneous scanning of transcripts from multiple cDNA samples and faster selection of differentially expressed transcripts of interest.

Differential Display Analysis of 2,3,7,8-Tetrachlorodibenzo-p-dioxin Identified Induction of Ras-related Nuclear Protein Binding Protein2 (RanBP2) Gene

  • Kim, Dong-Hak;Lim, Young-Ran;Park, Hyoung-Goo;Kim, Beom-Joon;Chun, Young-Jin
    • Toxicological Research
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    • v.25 no.1
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    • pp.35-40
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    • 2009
  • TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and related halogenated aromatic hydrocarbons elicit a diverse spectrum of biochemical and toxic responses in laboratory animals and mammalian cells in culture. Toxicity and carcinogenicity of TCDD is well established but the molecular mechanism is still poorly understood. Here, we found the noble responsive genes to TCDD using the differential display analysis. Treatment of HepG2 cells with TCDD showed a significantly different mRNA expression pattern from the untreated cells in differential display analysis. The differentially displayed bands were isolated and used as probes in dot blot and Northern blot analyses. Of thirty-five isolated differentially displayed bands, only two bands were confirmed as positive in dot blot and Northern blot analyses. The nucleotides sequences of these clones were analyzed and the search of Genebank database revealed that one clone is highly homologous with RanBP2 (Ras-related nuclear protein binding protein2; 92%) and the other is an unknown gene. RanBP2 is a nucleoporin with SUMO E3 ligase activity that functions in both nucleocytoplasmic transport and mitosis and its role as a novel tumor suppressor has been recently proposed. Thus, these results may suggest the clue elucidating the toxic mechanism of TCDD through RanBP2.

Development and Evaluation of Differential Pressure Type Mass Flow Controller for Semiconductor Fabrication Processing (반도체 공정용 차압식 질량 유량 제어 장치의 개발 및 성능 평가)

  • Ahn, Jin-Hong;Kang, Ki-Tai;Ahn, Kang-Ho
    • Journal of the Semiconductor & Display Technology
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    • v.7 no.3
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    • pp.29-34
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    • 2008
  • This paper describes the fabrication and characterization of a differential pressure type integrated mass-flow controller made of stainless steel for reactive and corrosive gases. The fabricated mass-flow controller is composed of a normally closed valve and differential pressure sensor. A stacked solenoid actuator mounted on a base-block is utilized for precise and rapid control of gas flow. The differential pressure flow sensor consisting of four diaphragms can detect a flow rate by deflection of diaphragm. By a feedback control from the flow sensor to the valve actuator, it is possible to keep the flow rate constant. This device shows a fast response less than 0.3 sec. Also, this device shows accuracy less than 0.1% of full scale. It is confirmed that this device is not attacked by toxic gas, so the integrated mass-flow controller can be applied to advanced semiconductor processes which need fine mass-flow control corrosive gases with fast response.

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Molecular Characterization of Seaweeds Using RAPD and Differential Display

  • HONG Yong-Ki;KIM Yong-Tae;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.770-778
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    • 1996
  • A rapid and economical method of simultaneous extraction of DNA and RNA from seaweeds has been developed by the use of lithium chloride. Lithium chloride facilitates the softening of cell walls resulting in a decrease in both compressive and tensile modulus of elasticity. The DNA was characterized by high molecular weight larger than 27 kb and a relative lack of carbohydrate and protein contamination. The DNA and RNA extracted by the method from many seaweeds were of sufficient quality to be used as a template for per amplification with a plant intergenic gene primer set, for RAPD analysis with arbitrary primers, and for differential display with arbitrary primers in the morphologically distinct regions of the matured Porphyra thallus. The cDNA polymorphism indicated that the reproductive tissue types (male, female, patch) had a relatively high degree of similarity; the vegetative tissue types (dividing, non-dividing) also showed a similar pattern with respect to each other. Holdfast tissue had very low similarity with the other tissues, but appeared most similar to vegetative non-dividing tissue type.

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Differential Display Detection of Acid-inducible Genes from Porphyra yezoensis Thalli (해조류 방사무늬김 (Porphra yezoensis) 엽체로부터 산 유도 유전자의 분리)

  • JIN Long-Guo;KANG Se-Eun;CHOI Jae-Suk;PARK Sun-Mee;PARK Jung-Youn;JIN Duck-Hee;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.4
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    • pp.269-274
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    • 2004
  • Genetic responses of the edible seaweed Porphyra yezoensis tissue to acid shock have been compared using differential display technique. The tissue was challenged in seawater containing $0.05{\%}$ hydrogen chloride (pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs. Total RNA extracted by the LiCl-guanidium method was reverse transcribed and amplified by PCR with arbitrary primers. The amplified fragment responded by the acid shock was selectively isolated from agarose gel and sequenced with DNA auto sequencer. Sequence (1056 bp) of the cDNA contained at least two genes for ASP7K (MW 7418) and ASP5K (MW 5512) proteins.

Analysis and Design Optimization of Interconnects for High-Speed LVDS Applications (고속 LVDS 응용을 위한 전송선 분석 및 설계 최적화)

  • Ryu, Jee-Youl;Noh, Seok-Ho
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.46 no.10
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    • pp.70-78
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    • 2009
  • This paper addresses the analysis and the design optimization of differential interconnects for high-speed Low-Voltage Differential Signaling (LVDS) applications. Thanks to the differential transmission and the low voltage swing, LVDS offers high data rates and improved noise immunity with significantly reduced power consumption in data communications, high-resolution display, and flat panel display. We present an improved model and new equations to reduce impedance mismatch and signal degradation in cascaded interconnects using optimization of interconnect design parameters such as trace width, trace height and trace space in differential printed circuit board (FPCB) transmission lines. We have carried out frequency-domain full-wave electromagnetic simulations, and time-domain transient simulations to evaluate the high-frequency characteristics of the differential FPCB interconnects. We believe that the proposed approach is very helpful to optimize high-speed differential FPCB interconnects for LVDS applications.

Design Optimization of Differential FPCB Transmission Line for Flat Panel Display Applications (평판디스플레이 응용을 위한 차동 FPCB 전송선 설계 최적화)

  • Ryu, Jee-Youl;Noh, Seok-Ho;Lee, Hyung-Joo
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.12 no.5
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    • pp.879-886
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    • 2008
  • This paper addresses the analysis and the design optimization of differential interconnects for Low-Voltage Differential Signaling (LVDS) applications. Thanks to the differential transmission and the low voltage swing, LVDS offers high data rates and improved noise immunity with significantly reduced power consumption in data communications, high-resolution display, and flat panel display. We present an improved model and new equations to reduce impedance mismatch and signal degradation in cascaded interconnects using optimization of interconnect design parameters such as trace width, trace height and trace space in differential flexible printed circuit board (FPCB) transmission lines. We have carried out frequency-domain full-wave electromagnetic simulations, time-domain transient simulations, and S-parameter simulations to evaluate the high-frequency characteristics of the differential FPCB interconnects. The 10% change in trace width produced change of approximately 6% and 5.6% in differential impedance for trace thickness of $17.5{\mu}m$ and $35{\mu}m$, respectively. The change in the trace space showed a little change. We believe that the proposed approach is very helpful to optimize high-speed differential FPCB interconnects for LVDS applications.

Involvement of Cytochrome c Oxidase Subunit I Gene during Neuronal Differentiation of PC12 Cells

  • Kang, Hyo-Jung;Chung, Jun-Mo;Lee, See-Woo
    • BMB Reports
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    • v.30 no.4
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    • pp.285-291
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    • 1997
  • It is becoming increasingly evident that significant changes in gene expression occur during the course of neuronal differentiation. Thus, it should be possible to gain information about the biochemical events by identifying differentially expressed genes in neuronal differentiation The PC12 cell line is a useful model system to investigate the molecular mechanism underlying neuronal differentiation and has been used extensively for the study of the molecular events that underlie the biological actions of nerve growth factor (NGF). In this study, we report an application of the recently described mRNA differential display method to analyze differential gene expression during neuronal differentiation. Using this technique, we have identified several cDNA tags expressed differentially during neuronal differentiation. Interestingly, one of these clones was cytochrome c oxidase subunit I (COX I) gene. The differential expression of COX I gene was confirmed by Northern blot analysis as well as RT-PCR. Southern blot analysis of the genomic DNA of PC12 cells revealed that COX I is a single gene. Induction of the oxidative enzyme might reflect the energy requirement in neuronal differentiation.

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Identification of Genes Induced by Low Temperature in Rice

  • Choi, Kyong-Hee;Choi, Hack-Sun;Lee, Choon-Hwan;Kwon, Young-Myung;Rhew, Tae-Hyong
    • BMB Reports
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    • v.30 no.4
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    • pp.292-295
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    • 1997
  • Exposure of seedling of rice (Oriza sativa cv.Dongin) to cold stress ($6^{circ}C$, 7day) induced differential gene expression. Differentially expressed polyadenylated RNA induced by low temperature were isolated and identified from the leaves of rice (Oriza sativa cv.Dongin) seedling by using the technique, differential display of reverse transcription through polymerase chain reaction (DDRT-PCR). Four bands of cDNAs were differentially displayed on the PAGE gel through DDRT-PCR, and among them three bands were those of overexpressed genes while one band was of an underexpressed gene One of the overexpressed cDNA was characterized. The size of the DDRT-PCR product was found to be about 200 bp. The sequence of the cloned DNA was compared with those of GenBank through a BLAST E-Mail server, and it was found to have no homologies in the nucleotide sequence with that of any known DNA: therefore, it was designated as RC101 The expression of the cold-stress induced-gene, RC101, was sustained with Northern Blot analysis by using the cloned DDRT-PCR product as a probe.

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Relationship between Differential Gene Expression in Ovary and Heterosis of Egg Number Traits in a Chicken Diallel Cross

  • Wang, Hui;Sun, Dong-Xiao;Yu, Ying;Wang, Dong;Zhang, Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.767-771
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    • 2005
  • In order to understand the molecular basis of chicken heterosis in reproduction traits, mRNA differential display (DDRT-PCR) methods were used to analyze the differential gene expression of ovary tissue between hybrids and their parental lines in a 4${\times}$4 diallel cross, involving 4 chicken breeds, which were White Plymouth Rock (E), CAU Brown (D), Silkies (C) and White Leghorn (A). Total of 331 differential displayed cDNA bands from 1,161 were displayed in the 4${\times}$4 diallel cross combinations with 30 pairs of primers, which shows the differences of gene expression between hybrids and their parental lines were very obvious in quantity and quality. Seven types of differential expression patterns were found: Co-dominance expressed pattern (T1), under-expression of parental fragments in hybrids (T2), over-expression of parental fragments in hybrids (T3), hybrid-absence expressed pattern (T4), single parentspecific expressed pattern (T5), dominant expression fragments of single parent in hybrids (T6), hybrid-specific expressed pattern (T7). Correlation analysis indicated that there were significant correlations between the pattern of T3 and the heterosis percentage of egg number of 32-week and 42-week old chickens(p<0.01), while there were negative significant correlations between the pattern of T7 and the heterosis percentage of egg number of 32-week and 42 week-old birds (p<0.01).