• Title/Summary/Keyword: Integration vector

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Field Performance and Morphological Characterization of Transgenic Codonopsis lanceolata Expressing $\gamma-TMT$ Gene.

  • Ghimire, Bimal Kumar;Li, Cheng Hao;Kil, Hyun-Young;Kim, Na-Young;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Chung, Ill-Min;Lee, Sun-Joo;Eom, Seok-Hyun;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.339-345
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    • 2007
  • Field performance and morphological characterization was conducted on seven transgenic lines of Codonopsis lanceolata expressing ${\gamma}-TMT$ gene. The shoots were obtained from leaf explants after co-cultivation with Agrobacterium tume-faciens strain LBA 4404 harboring a binary vector pYBI 121 that carried genes encoding ${\gamma}-Tocopherol$ methyltransferase gene (${\gamma}-TMT$) and a neomycin phosphotransferase II gene (npt II) for kanamycin resistance. The transgenic plants were transferred to a green house for acclimation. Integration of T-DNA into the $T_0\;and\;T_1$ generation of transgenic Codonopsis lanceolata genome was confirmed by the polymerase chain reaction and southern blot analysis. The progenies of transgenic plants showed phenotypic differences within the different lines and with relative to control plants. When grown in field, the transgenic plants in general exhibited increased fertility, significant improvement in the shoot weight, root weight, shoot height and rachis length with relation to the control plants. However, all seven independently derived transgenic lines produced normal flower with respect to its shape, size, color and seeds number at its maturity. Indicating that the addition of a selectable marker gene in the plant genome does not effect on seed germination and agronomic performance of transgenic Codonopsis lanceolata. $T_1$ progenies of these plants were obtained and evaluated together with control plant in a field experiment. Overall, the agronomic performance of $T_1$ progenies of transgenic Codonopsis lanceolata showed superior to that of the seed derived non-transgenic plant. In this study, we report on the morphological variation and agronomic performance of transgenic Codonopsis lanceolata developed by Agrobacterium transformation.

Integration of Motion Compensation Algorithm for Predictive Video Coding (예측 비디오 코딩을 위한 통합 움직임 보상 알고리즘)

  • Eum, Ho-Min;Park, Geun-Soo;Song, Moon-Ho
    • Journal of the Korean Institute of Telematics and Electronics S
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    • v.36S no.12
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    • pp.85-96
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    • 1999
  • In a number of predictive video compression standards, the motion is compensated by the block-based motion compensation (BMC). The effective motion field used for the prediction by the BMC is obviously discontinuous since one motion vector is used for the entire macro-block. The usage of discontinuous motion field for the prediction causes the blocky artifacts and one obvious approach for eliminating such artifacts is to use a smoothed motion field. The optimal procedure will depend on the type of motion within the video. In this paper, several procedures for the motion vectors are considered. For any interpolation or approaches, however, the motion vectors as provided by the block matching algorithm(BMA) are no longer optimal. The optimum motion vectors(still one per macro-block) must minimize the of the displaced frame difference (DFD). We propose a unified algorithm that computes the optimum motion vectors to minimize the of the DFD using a conjugate gradient search. The proposed algorithm has been implemented and tested for the affine transformation based motion compensation (ATMC), the bilinear transformation based motion compensation (BTMC) and our own filtered motion compensation(FMC). The performance of these different approaches will be compared against the BMC.

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EST Analysis system for panning gene

  • Hur, Cheol-Goo;Lim, So-Hyung;Goh, Sung-Ho;Shin, Min-Su;Cho, Hwan-Gue
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.21-22
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    • 2000
  • Expressed sequence tags (EFTs) are the partial segments of cDNA produced from 5 or 3 single-pass sequencing of cDNA clones, error-prone and generated in highly redundant sets. Advancement and expansion of Genomics made biologists to generate huge amount of ESTs from variety of organisms-human, microorganisms as well as plants, and the cumulated number of ESTs is over 5.3 million, As the EST data being accumulate more rapidly, it becomes bigger that the needs of the EST analysis tools for extraction of biological meaning from EST data. Among the several needs of EST analyses, the extraction of protein sequence or functional motifs from ESTs are important for the identification of their function in vivo. To accomplish that purpose the precise and accurate identification of the region where the coding sequences (CDSs) is a crucial problem to solve primarily, and it will be helpful to extract and detect of genuine CD5s and protein motifs from EST collections. Although several public tools are available for EST analysis, there is not any one to accomplish the object. Furthermore, they are not targeted to the plant ESTs but human or microorganism. Thus, to correspond the urgent needs of collaborators deals with plant ESTs and to establish the analysis system to be used as general-purpose public software we constructed the pipelined-EST analysis system by integration of public software components. The software we used are as follows - Phred/Cross-match for the quality control and vector screening, NCBI Blast for the similarity searching, ICATools for the EST clustering, Phrap for EST contig assembly, and BLOCKS/Prosite for protein motif searching. The sample data set used for the construction and verification of this system was 1,386 ESTs from human intrathymic T-cells that verified using UniGene and Nr database of NCBI. The approach for the extraction of CDSs from sample data set was carried out by comparison between sample data and protein sequences/motif database, determining matched protein sequences/motifs that agree with our defined parameters, and extracting the regions that shows similarities. In recent future, in addition to these components, it is supposed to be also integrated into our system and served that the software for the peptide mass spectrometry fingerprint analysis, one of the proteomics fields. This pipelined-EST analysis system will extend our knowledge on the plant ESTs and proteins by identification of unknown-genes.

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The Overexpression of Subtilisin Enzyme Using Mutations on Transition State Regulatory Proteins of AprE Promoter and Development of Bacillus subtilis Host System (AprE Promoter전이상태 조절인자 변이주를 이용한 공업적 효소의 과발현과 고초균 숙주계의 개발)

  • 류성호;박승환김병기
    • KSBB Journal
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    • v.11 no.1
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    • pp.8-14
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    • 1996
  • Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

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Transformation of the Diatom Phaeodactylum tricornutum with its Endogenous (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase Gene (Phaeodactylum tricornutum의 (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase 유전자의 형질전환)

  • Shin, Bok-Kyu;Jung, Yu-Jin;Kim, Sang-Min;Pan, Cheol-Ho
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.273-279
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    • 2015
  • Phaeodactylum tricornutum is a model diatom that its genomic information and biological tools are well established. In this study, a gene encoding (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (PtHDR), a terminal enzyme of the methylerythritol phosphate pathway regulating chlorophyll and carotenoid biosynthesis, was isolated from P. tricornutum. The isolated gene was cloned into pPha-T1 vector containing fcpA promoter to prepare pPha-T1-HDR plasmid. As a positive control, pPha-T1-eGFP plasmid was constructed with egfp gene. Stable nuclear transformation was carried out with these plasmids by particle bombardment method and zeocin resistant colonies of P. tricornutum were selected on f/2 agar plate. In result, transformation efficiency was evaluated according to the amount of plasmid DNA coated with gold particles. Integration of introduced plasmids was confirmed with genomic DNA of each transformant by polymerase chain reaction. The eGFP fluorescence was visible in the cytoplasm, indicating that eGFP was successively expressed in P. tricornutum system. The transcript level of exogenous Pthdr gene was evaluated with the obtained transformants. The results presented here demonstrated that introduction of Pthdr gene into P. tricornutum chromosome succeeded and expression of PtHDR was enhanced under the fcpA promoter.

P Element-Mediated Transformation with the rosy Gene in Drosophila melanogaster (D. melanogaster에 있어서 P Element를 이용한 rosy 유전자의 형질전환)

  • Kim, Wook;Kidwell, Margaret G.
    • The Korean Journal of Zoology
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    • v.38 no.3
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    • pp.340-347
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    • 1995
  • We have used two kinds of P element constructs, Pc[(ry+)B] and p[(ry+)$\Delta$SX9], for genetic transformation by microinjection of D. melanogaster. Pc[(ry+)B] construct carrying the rosy gene within an autonomous P element was injected into a true M strain caring the ry506. mutation. The source of transposase for microinjection and transformation was provided by a P element helper plasmid designated p-$\Delta$2-3hs$\pi$, which was co-injected with nonautonomous P[(ry+)$\Delta$SX9] construct into same ry506 M strains. A dechorination method was adopted and 35 independent transformed lines were obtained froin 1143 G0 Injected (35/1143). About 20% of the injected embryos eclosed as adults. Among G0 eclosed flies, approximately 40% exhibited eye color that was similar to wild-type (ry+), but about 60% of fertile G0 transformed lines appeared to have no G1 transformants. Therefore it is unlikely that G0 expression requires integration of the rosy transposon into chromosomes. Pc[(ry+)B] and P[(ry+)$\Delta$SX9] constructs were found to be nearly same in the frequency of element-mediated transformation. On the basis of these results, nonautonomous P elements constructs could he used as same effective vectors in P element-mediated transformation for introducing and fixing genes in insect populations.

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Predicting Stock Liquidity by Using Ensemble Data Mining Methods

  • Bae, Eun Chan;Lee, Kun Chang
    • Journal of the Korea Society of Computer and Information
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    • v.21 no.6
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    • pp.9-19
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    • 2016
  • In finance literature, stock liquidity showing how stocks can be cashed out in the market has received rich attentions from both academicians and practitioners. The reasons are plenty. First, it is known that stock liquidity affects significantly asset pricing. Second, macroeconomic announcements influence liquidity in the stock market. Therefore, stock liquidity itself affects investors' decision and managers' decision as well. Though there exist a great deal of literature about stock liquidity in finance literature, it is quite clear that there are no studies attempting to investigate the stock liquidity issue as one of decision making problems. In finance literature, most of stock liquidity studies had dealt with limited views such as how much it influences stock price, which variables are associated with describing the stock liquidity significantly, etc. However, this paper posits that stock liquidity issue may become a serious decision-making problem, and then be handled by using data mining techniques to estimate its future extent with statistical validity. In this sense, we collected financial data set from a number of manufacturing companies listed in KRX (Korea Exchange) during the period of 2010 to 2013. The reason why we selected dataset from 2010 was to avoid the after-shocks of financial crisis that occurred in 2008. We used Fn-GuidPro system to gather total 5,700 financial data set. Stock liquidity measure was computed by the procedures proposed by Amihud (2002) which is known to show best metrics for showing relationship with daily return. We applied five data mining techniques (or classifiers) such as Bayesian network, support vector machine (SVM), decision tree, neural network, and ensemble method. Bayesian networks include GBN (General Bayesian Network), NBN (Naive BN), TAN (Tree Augmented NBN). Decision tree uses CART and C4.5. Regression result was used as a benchmarking performance. Ensemble method uses two types-integration of two classifiers, and three classifiers. Ensemble method is based on voting for the sake of integrating classifiers. Among the single classifiers, CART showed best performance with 48.2%, compared with 37.18% by regression. Among the ensemble methods, the result from integrating TAN, CART, and SVM was best with 49.25%. Through the additional analysis in individual industries, those relatively stabilized industries like electronic appliances, wholesale & retailing, woods, leather-bags-shoes showed better performance over 50%.

Developing a Gene-trapping Approach for Gene Identification Using Nuclear Transfer in Zebrafish (지브라물고기 복제방법에 의한 유전자 동정 및 유전자트랩법 개발)

  • Lee, K.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.155-164
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    • 2004
  • This involves identifying and cloning trapped genes from cultured cells carrying the gene-trap constructs and generating cloned zebrafish using these cells for functional study. Gene-trapping studies in gene-trapped cells were carried out in initial and cloned zebrafish carrying gene-trap events were successfully produced based on the nuclear transplantation technique. Two kind of retroviral gene-trap constructs were adopted. The first one(SA/GFP-TP), constructed in my laboratory, carries a GFP reporter gene containing a splicing acceptor and an internal neo gene. The second one(Neo-TP), obtained from Dr. Hicks (Hicks et al., 1997), contains a promoter-less neo gene located in the LTR sequence of a retroviral vector. The infected cells were subjected to drug selection(neomycin treatment) because the two constructs carry the neomycin resistant gene. All those cells survived the neomycin treatment should carry the proviral insertions. For Neo-TP, Isolated DNA from the neomycin-resistant fibroblast cells infected by Neo-TP, was digested with EcoR1 restriction enzyme and transformed into bacteria after ligation. This procedure led to the isolation of seven clones carrying flanking cellular DNA with a typical retroviral integration signature sequence. These clones contained genomic DNA ranging from 1kb to 7kb and sequences of 300-600 bp were obtained from each of the rescued plasmids. Database searching showed that all of them share high homology to zebrafish sequences. For fish cloning using tagged cells, initially, nucleus donors directly selected from a mixture of cells(Neo-TP cells) were used. A total of 44 embryos(3.7%) out of 1179 transplants were reached blastula stage; 8 of these embryos(0.8%) hatched and 3(0.3%) of them survived to adulthood. One out of three lived cloned zebrafish has an amplified fragment and was labeled with 32P.

Design and Implementation of Co-Verification Environments based-on SystemVerilog & SystemC (SystemVerilog와 SystemC 기반의 통합검증환경 설계 및 구현)

  • You, Myoung-Keun;Song, Gi-Yong
    • Journal of the Institute of Convergence Signal Processing
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    • v.10 no.4
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    • pp.274-279
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    • 2009
  • The flow of a universal system-level design methodology consists of system specification, system-level hardware/software partitioning, co-design, co-verification using virtual or physical prototype, and system integration. In this paper, verification environments based-on SystemVerilog and SystemC, one is native-code co-verification environment which makes prompt functional verification possible and another is SystemVerilog layered testbench which makes clock-level verification possible, are implemented. In native-code co-verification, HW and SW parts of SoC are respectively designed with SystemVerilog and SystemC after HW/SW partitioning using SystemC, then the functional interaction between HW and SW parts is carried out as one simulation process. SystemVerilog layered testbench is a verification environment including corner case test of DUT through the randomly generated test-vector. We adopt SystemC to design a component of verification environment which has multiple inheritance, and we combine SystemC design unit with the SystemVerilog layered testbench using SystemVerilog DPI and ModelSim macro. As multiple inheritance is useful for creating class types that combine the properties of two or more class types, the design of verification environment adopting SystemC in this paper can increase the code reusability.

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Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.