• Title/Summary/Keyword: Hybrid Library

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Isolation of CONSTANS as a TGA4/OBF4 Interacting Protein

  • Song, Young Hun;Song, Na Young;Shin, Su Young;Kim, Hye Jin;Yun, Dae-Jin;Lim, Chae Oh;Lee, Sang Yeol;Kang, Kyu Young;Hong, Jong Chan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.559-565
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    • 2008
  • Members of the TGA family of basic domain/leucine zipper transcription factors regulate defense genes through physical interaction with NON-EXPRESSOR OF PR1 (NPR1). Of the seven TGA family members, TGA4/octopine synthase (ocs)-element-binding factor 4 (OBF4) is the least understood. Here we present evidence for a novel function of OBF4 as a regulator of flowering. We identified CONSTANS (CO), a positive regulator of floral induction, as an OBF4-interacting protein, in a yeast two-hybrid library screen. OBF4 interacts with the B-box region of CO. The abundance of OBF4 mRNA cycles with a 24 h rhythm under both long-day (LD) and short-day (SD) conditions, with significantly higher levels during the night than during the day. Electrophoretic mobility shift assays revealed that OBF4 binds to the promoter of the FLOWERING LOCUS T (FT) gene, a direct target of CO. We also found that, like CO and FT, an OBF4:GUS construct was prominently expressed in the vascular tissues of leaf, indicating that OBF4 can regulate FT expression through the formation of a protein complex with CO. Taken together, our results suggest that OBF4 may act as a link between defense responses and flowering.

Functional Genomics for Mass Analysis of Useful Genes in Panax ginseng C.A. Meyer (인삼의 유용유전자원 확보를 위한 기능 유전체연구)

  • Yang, Deok-Chun
    • Proceedings of the Ginseng society Conference
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    • 2004.05a
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    • pp.17-28
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    • 2004
  • As Korean ginseng is hybrid, an individual variation is very severe, and it takes long times in new breeding because it is required 4 years to pick the seed. But, transformation technique makes the high-functional breeding in short time. The focus of these ginseng studies is to find and secure the useful gene. And it is urgent to accumulate the fundamental data for the molecular breeding and secure the useful genes. Therefore, transformation and soil acclimatization technique are necessary to molecular breeding in use of the introduction of functional genes. In this study, it add to secure of new regulation gene and useful gene as to accumulate the fundamental data for the place where it will contribute to raise the national competitive power. To analyze the useful genes in large scale, we constructed CDNA libraries with various tissues, species, and treated tissue. EST analysis of ginseng perform in large scale and build the EST database of ginseng. We perform the full length sequencing about the selected lots of clones that include the entire open reading frame of the amino acid residues and construct cDNA chip with the parental EST clones. Establishment of the transformation and a soil acclimatization system throuth the re-introduction of the selected ginseng gene that related with the secondary metabolism and anti-stress into the ginseng.

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PRIP, a Novel Ins(1,4,5)P3 Binding Protein, Functional Significance in Ca2+ Signaling and Extension to Neuroscience and Beyond

  • Kanematsu, Takashi;Takeuchi, Hiroshi;Terunuma, Miho;Hirata, Masato
    • Molecules and Cells
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    • v.20 no.3
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    • pp.305-314
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    • 2005
  • Investigation of chemically synthesized inositol 1,4,5-trisphosphate [$Ins(1,4,5)P_3$] analogs has led to the isolation of a novel binding protein with a molecular size of 130 kDa, characterized as a molecule with similar domain organization to phospholipase C-${\delta}1$ (PLC-${\delta}1$) but lacking the enzymatic activity. An isoform of the molecule was subsequently identified, and these molecules have been named PRIP (PLC-related, but catalytically inactive protein), with the two isoforms named PRIP-1 and -2. Regarding its ability to bind $Ins(1,4,5)P_3$ via the pleckstrin homology domain, the involvement of PRIP-1 in $Ins(1,4,5)P_3$-mediated $Ca^{2+}$ signaling was examined using COS-1 cells overexpressing PRIP-1 and cultured neurons prepared from PRIP-1 knock-out mice. Yeast two hybrid screening of a brain cDNA library using a unique N-terminus as bait identified GABARAP ($GABA_A$ receptor associated protein) and PP1 (protein phosphatase 1), which led us to examine the possible involvement of PRIP in $GABA_A$ receptor signaling. For this purpose PRIP knock-out mice were analyzed for $GABA_A$ receptor function in relation to the action of benzodiazepines from the electrophysiological and behavioral aspects. During the course of these experiments we found that PRIP also binds to the b-subunit of $GABA_A$ receptors and PP2A (protein phosphtase 2A). Here, we summarize how PRIP is involved in $Ins(1,4,5)P_3$-mediated $Ca^{2+}$ signaling and $GABA_A$ receptor signaling based on the characteristics of binding molecules.

A single-memory based FFT/IFFT core generator for OFDM modulation/demodulation (OFDM 변복조를 위한 단일 메모리 구조의 FFT/IFFT 코어 생성기)

  • Yeem, Chang-Wan;Jeon, Heung-Woo;Shin, Kyung-Wook
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2009.05a
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    • pp.253-256
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    • 2009
  • This paper describes a core generator (FFT_Core_Gen) which generates Verilog HDL models of 8 different FFT/IFFT cores with $N=64{\times}2^k$($0{\leq}k{\leq}7$ for OFDM-based communication systems. The generated FFT/IFFT cores are based on in-place single memory architecture, and use a hybrid structure of radix-4 and radix-2 DIF algorithm to accommodate various FFT lengths. To achieve both memory reduction and the improved SQNR, a conditional scaling technique is adopted, which conditionally scales the intermediate results of each computational stage, and the internal data and twiddle factor has 14 bits. The generated FFT/IFFT cores have the SQNR of 58-dB for N=8,192 and 63-dB for N=64. The cores synthesized with a $0.35-{\mu}m$ CMOS standard cell library can operate with 75-MHz@3.3-V, and a 8,192-point FFT can be computed in $762.7-{\mu}s$, thus the cores satisfy the specifications of wireless LAN, DMB, and DVB systems.

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Control of Trophoblast Gene Expression and Cell Differentiation

  • Cheon, Jong-Yun
    • 대한생식의학회:학술대회논문집
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    • 2001.03a
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    • pp.195-205
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    • 2001
  • 태반 영양배엽 (trophoblast)은 포유동물의 발생과정 중 가장 먼저 분화되는 세포로서, 자궁환경내에서 배아가 착상, 발생, 및 분화하기 위해서 반드시 필요한 태반을 형성하는 색심적인 세포이다. 영양배엽 세포의 분화과정중의 결함은 배아의 사산이나 임신질환 등의 치명적 결과를 초래한다. 하지만, 영양배엽 세포의 분화를 조절하는 분자생물학적인 메카니즘은 아직 규명되지 않고 있다. 영양배엽 세포의 분화를 조절하는 경로를 규경하기 위한 선결과제는 분화된 영양배엽 세포에서만 발현하는 많은 유전자들이 밝혀져야만 한다. 본 연구팀은 최근에 분화된 영양배엽 세포에서만 발현하는 두 종류의 새로운 유전자들을 찾았다. 한 종류는 homeobox를 보유하고 있는 조절 유전자 Psx이고, 다른 한 종류는 임신호르몬인 태반 프로락틴 라이크 단백질 유전자 PLP-C${\beta}$이다. 본 연구과제의 목표는 이들 유전자의 기능과 조절 메카니즘을 규명함으로써, 영양배엽 세포의 분화를 조절하는 조절경로를 밝히는 것이다. 이를 위하여 다음과 같은 일련의 연구를 수행할 것이다. 1) Psx 유전자가 분화된 영양배엽 세포에서만 발현케 하는 조절 메카니즘을 규명하기 위해 functional assays, in vitro footprinting, gel mobility shift assays, 생쥐형질전화, UV crosslinking, Southwestern blot 등의 방법을 통해 Psx 유전자의 cis-acting 요인과 trans-acting factor를 밝혀 분석한다. 2) 영양배엽 세포의 분화조절 경로를 규명하기 위해 random oligonuclotide library screening, DD-PCR, subtractive screening 등의 방법을 이용하여 Psx 유전자에 의해 조절되는 하부유전자를 밝힌다. 3) Psx 유전자를 knock-out시켜 영양배엽 세포가 발달 및 분화하는데 미치는 역할을 밝힌다. 4) Yeast two-hybrid screening방법을 이용하여 태반 프로락틴 유전자의 수용체를 찾아 이들의 신호전달 기전을 밝힌다. 제1차년 연구결과로서, mouse와 rat으로부터 각각 Psx 유전자의 genomic DNA를 클로닝하여, 유전자 구조를 비교한 결과, mouse Psx (mPsx2)는 4개의 exons으로 이루어져 있는 반면에, rat Psx (Psx3)는 3개의 exons으로 구성되어 있었다. 즉, rPsx3는 mPsx2의 exon1이 없었다. Notrhern blot과 in situ hybridization 분석에 의해 mouse와 rat에서 Psx 유전자가 다르게 발현 조절되는 현상을 밝혔다. 실제로 mPsx2와 rPsx3의 5'-flanking지역을 클로닝하여 염기서열 분석 결과 전혀 homology를 찾을 수 없었다. 또한, 이들 각각 promoter의 activity를 luciferase reporter를 이용하여 조사한 결과 Rcho-1 trophoblast cells에서 각기 다른 activity를 보여 주는 것을 발견하였다. Psx 유전자의 transcription start sites는 Primer extension에 의해 밝혔다. 또한 Psx2 유전자를 knock-out 시키기 위해 targeting vector를 Osdupde1에 제작하였다. 본 과제를 시작할 때 새로운 프로락틴 유전자 하나를 클로닝하여 이 유전자를 PLP-I라고 이름을 붙였다. 이 후 이 유전자 (PLP-I)는 PLP-C${\beta}$라고 이름을 붙이게 되었다. Mouse PLP-C${\beta}$ 유전자의 counterpart를 rat에서 찾아 염기서열을 비교한 결과 mouse와 rat에서 PLP-C${\beta}$유전자의 homology는 약 79% (amino acid level)였다. 본 연구과정을 통해 또 하나의 새로운 PLP-C subfamily member를 mouse로부터 클로닝 하였고, 이 유전자를 PLP-C${\gamma}$라 하였다. PLP-C${\beta}$와 PLP-C${\gamma}$의 발현 유형은 Northern blot과 in 냐셔 hybridization 분석에 의해 태반의 제한된 spongitrophoblast와 trophoblast giant cells에서만 발현하는 것을 밝혔다. 놀랍게도 이들 두 새로운 유전자는 alternative splicing에 의해 두 종류의 isoform이 있음을 밝혔다. PLP family member 유전자로서 splicing에 의한 isoforms을 보여 주는 유전자로는 PLP-C${\beta}$와 PLP-C${\gamma}$가 최초이다. 이들 isoform mRNAs의 발현 유형은 RT-PCR 방법을 이용하여 규명하였다. 또 하나의 새로운 발견은 PLP-C${\beta}$와 PLP-C${\gamma}$가 독특한 유전자 구조를 갖고 있었다. 즉, PLP-C${\beta}$는 exon3의 alternative splicing에 의해 5개 혹은 6개의 exons을 갖는 two isoforms이 생긴다. 반면에 PLP-C${\gamma}$는 exon2가 alternative splcing이 되면서 7개의 exons을 갖거나 6개의 exons을 갖는 isoforms을 만든다. 그리고, PLP-C${\gamma}$의 promoter activity를 trophoblast Rcho-l${\gamma}$ 세포주를 이용하여 PLP-C${\gamma}$ 의 1.5 kb 5'-flanking 지역이 trophoblast-specific promoter activity를 갖고 있음을 밝혔다. PLP-C${\gamma}$ 유전자의 transcription start site는 Primer extension에 의해 밝혔다. 제 1차 년도의 연구결과를 토대로, 2차년에서는 다음단계의 연구를 수행하고자 한다. 즉, 1) mPsx2와 rPsx3의 promoter를 비교분석 함으로서 mouse와 rat에서 Psx 유전자가 다르게 조절되는 메카니즘 규명, 2) Psx와 PLP-C 유전자의 promoter에 있는 cis-acting elements 탐색, 3) Psx2와 Psx3의 단백질을 이용하여 이들이 binding하는 target sequence 규명, 4) 제작한 Psx2 targeting vector를 이용하여 ES cells에서 Psx2 유전자 knock-out, 5) Psx 유전자를 과발현시키는 세포주를 만들고 Psx에 의해 조절되는 유전자 탐색, 6) 새로 밝히 PLP-C members 유전자들의 조절기전을 Rcho-1 세포주를 이용하여 여러 거지 성장인자와 다른 호르몬에 대한 반응을 탐색, 7) Psx와 PLP-C${\gamma}$ 유전자의 chromosomal mapping 등을 밝힐 것이다.

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Emoticon by Emotions: The Development of an Emoticon Recommendation System Based on Consumer Emotions (Emoticon by Emotions: 소비자 감성 기반 이모티콘 추천 시스템 개발)

  • Kim, Keon-Woo;Park, Do-Hyung
    • Journal of Intelligence and Information Systems
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    • v.24 no.1
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    • pp.227-252
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    • 2018
  • The evolution of instant communication has mirrored the development of the Internet and messenger applications are among the most representative manifestations of instant communication technologies. In messenger applications, senders use emoticons to supplement the emotions conveyed in the text of their messages. The fact that communication via messenger applications is not face-to-face makes it difficult for senders to communicate their emotions to message recipients. Emoticons have long been used as symbols that indicate the moods of speakers. However, at present, emoticon-use is evolving into a means of conveying the psychological states of consumers who want to express individual characteristics and personality quirks while communicating their emotions to others. The fact that companies like KakaoTalk, Line, Apple, etc. have begun conducting emoticon business and sales of related content are expected to gradually increase testifies to the significance of this phenomenon. Nevertheless, despite the development of emoticons themselves and the growth of the emoticon market, no suitable emoticon recommendation system has yet been developed. Even KakaoTalk, a messenger application that commands more than 90% of domestic market share in South Korea, just grouped in to popularity, most recent, or brief category. This means consumers face the inconvenience of constantly scrolling around to locate the emoticons they want. The creation of an emoticon recommendation system would improve consumer convenience and satisfaction and increase the sales revenue of companies the sell emoticons. To recommend appropriate emoticons, it is necessary to quantify the emotions that the consumer sees and emotions. Such quantification will enable us to analyze the characteristics and emotions felt by consumers who used similar emoticons, which, in turn, will facilitate our emoticon recommendations for consumers. One way to quantify emoticons use is metadata-ization. Metadata-ization is a means of structuring or organizing unstructured and semi-structured data to extract meaning. By structuring unstructured emoticon data through metadata-ization, we can easily classify emoticons based on the emotions consumers want to express. To determine emoticons' precise emotions, we had to consider sub-detail expressions-not only the seven common emotional adjectives but also the metaphorical expressions that appear only in South Korean proved by previous studies related to emotion focusing on the emoticon's characteristics. We therefore collected the sub-detail expressions of emotion based on the "Shape", "Color" and "Adumbration". Moreover, to design a highly accurate recommendation system, we considered both emotion-technical indexes and emoticon-emotional indexes. We then identified 14 features of emoticon-technical indexes and selected 36 emotional adjectives. The 36 emotional adjectives consisted of contrasting adjectives, which we reduced to 18, and we measured the 18 emotional adjectives using 40 emoticon sets randomly selected from the top-ranked emoticons in the KakaoTalk shop. We surveyed 277 consumers in their mid-twenties who had experience purchasing emoticons; we recruited them online and asked them to evaluate five different emoticon sets. After data acquisition, we conducted a factor analysis of emoticon-emotional factors. We extracted four factors that we named "Comic", Softness", "Modernity" and "Transparency". We analyzed both the relationship between indexes and consumer attitude and the relationship between emoticon-technical indexes and emoticon-emotional factors. Through this process, we confirmed that the emoticon-technical indexes did not directly affect consumer attitudes but had a mediating effect on consumer attitudes through emoticon-emotional factors. The results of the analysis revealed the mechanism consumers use to evaluate emoticons; the results also showed that consumers' emoticon-technical indexes affected emoticon-emotional factors and that the emoticon-emotional factors affected consumer satisfaction. We therefore designed the emoticon recommendation system using only four emoticon-emotional factors; we created a recommendation method to calculate the Euclidean distance from each factors' emotion. In an attempt to increase the accuracy of the emoticon recommendation system, we compared the emotional patterns of selected emoticons with the recommended emoticons. The emotional patterns corresponded in principle. We verified the emoticon recommendation system by testing prediction accuracy; the predictions were 81.02% accurate in the first result, 76.64% accurate in the second, and 81.63% accurate in the third. This study developed a methodology that can be used in various fields academically and practically. We expect that the novel emoticon recommendation system we designed will increase emoticon sales for companies who conduct business in this domain and make consumer experiences more convenient. In addition, this study served as an important first step in the development of an intelligent emoticon recommendation system. The emotional factors proposed in this study could be collected in an emotional library that could serve as an emotion index for evaluation when new emoticons are released. Moreover, by combining the accumulated emotional library with company sales data, sales information, and consumer data, companies could develop hybrid recommendation systems that would bolster convenience for consumers and serve as intellectual assets that companies could strategically deploy.