• Title/Summary/Keyword: Double sequences

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Recombination Activating Gene 1 Product Alone Possesses Endonucleolytic Activity

  • Kim, Deok-Ryong
    • BMB Reports
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    • v.36 no.2
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    • pp.201-206
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    • 2003
  • Two lymphoid-specific proteins, RAG1 and RAG2, are required for the initiation of the V(D)J recombination in vitro. The V(D)J cleavage that is mediated by RAG proteins at the border between the coding and signal sequences results in the production of a hairpin at the coding end and a double-stranded break at the signal end. Two hairpin coding ends are re-opened, modified, and sealed; whereas, the signal ends are directly ligated. Here I report that only RAG1 can carry out a distinct endonucleolytic activity in vitro using an oligonucleotide substrate that is tethered by a short single-stranded DNA. The purified RAG1 protein alone formed a nick at the near position to the recombination signal sequence. This endonucleolytic activity was eliminated by immunoprecipitation using the RAG1-specific antibody, and required the 3'-hydroxy group. All of the RAG1 mutants that were incapable of the nick and hairpin formation in the V(D)J cleavage analysis also showed this new endonucleolytic activity. This suggests that the nicking activity that was observed might be functionally different from the nick formation in the V(D)J cleavage.

An efficient gene targeting system using homologous recombination in plants (식물에서의 상동재조합을 이용한 효율적인 진타겟팅 시스템)

  • Kwon, Yong-Ik;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.154-160
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    • 2015
  • The plant breeding technology was developed with genetic engineering. Many researchers and breeders have turned from traditional breeding to molecular breeding. Genetically modified organisms (GMO) were developed via molecular breeding technology. Currently, molecular breeding technologies facilitate efficient plant breeding without introducing foreign genes, in virtue by of gene editing technology. Gene targeting (GT) via homologous recombination (HR) is one of the best gene editing methods available to modify specific DNA sequences in genomes. GT utilizes DNA repair pathways. Thus, DNA repair systems are controlled to enhance HR processing. Engineered sequence specific endonucleases were applied to improve GT efficiency. Engineered sequence specific endonucleases like the zinc finger nuclease (ZFN), TAL effector nuclease (TALEN), and CRISPR-Cas9 create DNA double-strand breaks (DSB) that can stimulate HR at a target site. RecQl4, Exo1 and Rad51 are effectors that enhance DSB repair via the HR pathway. This review focuses on recent developments in engineered sequence specific endonucleases and ways to improve the efficiency of GT via HR effectors in plants.

New Hairpin RNAi Vector with Brassica rapa ssp. pekinensis Intron for Gene Silencing in Plants

  • Lee, Gi-Ho;Lee, Gang-Seob;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.323-332
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    • 2017
  • Homology-specific transcriptional and post-transcriptional silencing, an intrinsic mechanism of gene regulation in most eukaryotes, can be induced by anti-sense, co-suppression, or hairpin-based double-stranded RNA. Hairpin-based RNA interference (RNAi) has been applied to analyze gene function and genetically modify crops. However, RNAi vector construction usually requires high-cost cloning steps and large amounts of time, or involves methods that are protected by intellectual property rights. We describe a more effective method for generating intron-spliced RNAi constructs. To produce intron-spliced hairpin RNA, an RNAi cassette was ligated with the first intron and splicing sequences of the Brassica rapa ssp. pekinensis histone deacetylase 1 gene. This method requires a single ligation of the PCR-amplified target gene to SpeI-NcoI and SacI-BglII enzyme sites to create a gene-specific silencing construct. We named the resulting binary vector system pKHi and verified its functionality by constructing a vector to silence DIHYDROFLAVONOL 4-REDUCTASE (DFR), transforming it into tobacco plants, and confirming DFR gene-silencing via PCR, RT-qPCR, and analysis of the accumulation of small interfering RNAs. Reduction of anthocyanin biosynthesis was also confirmed by analyzing flower color of the transgenic tobacco plants. This study demonstrates that small interfering RNAs generated through the pKHi vector system can efficiently silence target genes and could be used in developing genetically modified crops.

DNA Ligand - Redox Active Molecule Conjugates as an Electrochemical DNA Probe

  • Ihara, Toshihiro;Maruo, Voshiyuki;Uto, Yoshihiro;Takenaka, Shigeori;Takagi, Makoto
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.887-894
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    • 1995
  • Toward the development of universal, sensitive, and convenient method of DNA (or RNA) detection, two kinds of electrochemically active DNA ligands. acridine - viologen and oligonucleotide - ferrocene conjugate, were prepared. Thermodynamic and electrochemical study revealed that these probes bound strongly to DNA, and showed a typical cyclic voltammograms, indicating a potential for use as a reversible electrochemical labelling agent for DNA. Especially, using the electrochemically active oligonucleotide, we have been able to demonstrate the detection of DNA at femtomole levels by HPLC equipped with ordinary electrochemical detector (ECD). These results lead to the conclusion that the redox-active probes are very useful for the microanalysis of nucleic acid due to the stabilily of the complexes, high detection sensitivity, and wide applicability to the target structures (single- and double strands) and sequences.

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nif-Gene Organization and Nucleotide Sequence of nifV, nifH, D, K and nifE from Frankia Strain FaCl

  • An, Chung-Sun
    • Proceedings of the Zoological Society Korea Conference
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    • 1995.10b
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    • pp.120-120
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    • 1995
  • The total size of the pF AR1, a genomic clone of Frankia FaCI, was estimated to be about 44Kb by summation of the individual fragment length generated by single or double restriction enzymes. Southern hybridization analyses with Azotobacter vinelandii nif-genes as probes and partial sequencing analyses of the subclones revealed that organization of the nif-gene in the FaCI strain was nifV, H, D, K, E, N, X, W, B. The organization of the structural genes for nitrogenase is the same in this Frankia strain as it is in most other nitrogen-fixing prokaryotes but the positioning of the nifV-like gene relative to the nifHDK cluster differs. A consensus nif-promoter-like sequence, found at 5' of nifH, was not detected upstream of the niJV-like gene. nifV-like gene contained a ORF of 1206 NT encoding 401 amino acids. The nucleotide sequence and deduced amino acid sequence of the gene exhibit homology value of 65% and 41% with that from A vinelandii, respectively. The putative Shine-Dargamo sequences were present preceding nitK, nifH, D, K, and nifE, and in nitK gene putative start codon GTG was detected instead of A TG. The nucleotide and amino acid sequence of niIK of FaCI showed 82% and 76% homolgy with those of Frankia HFPCc 13, respectively. Amino acid sequence of niIK showed 69% and 61% homology with those of A vinelandii, Klebsiella pnewnoniae, respectively, while that of nifE 73% and 71%, respecti vely.i vely.

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Segmentation of Moving Multiple Vehicles using Logic Operations (논리연산을 이용한 주행차량 영상분할)

  • Choi Kiho
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.1 no.1
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    • pp.10-16
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    • 2002
  • In this paper, a novel algorithm for segmentation of moving multiple vehicles in video sequences using logic operations is proposed. For the case of multiple vehicles in a scene, the proposed algorithm begins with a robust double-edge image derived from the difference between two successive frames using exclusive OR operation. After extracting only the edges of moving multiple vehicles using Laplacian filter, AND operation and dilation operation, the image is segmented into moving multiple vehicle image. The features of moving vehicles can be directly extracted from the segmented images. The proposed algorithm has no the two preprocessing steps, so it can reduce noises which are norm at in preprocessing of the original images. The algorithm is more simplified using logic operations. The proposed algorithm is evaluated on an outdoor video sequence with moving multiple vehicles in 90,000 frames of 30fps by a low-end video camera and produces promising results.

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Performance Comparison of HEVC and H.264/AVC Standards in Broadcasting Environments

  • Dissanayake, Maheshi B.;Abeyrathna, Dilanga L.B.
    • Journal of Information Processing Systems
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    • v.11 no.3
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    • pp.483-494
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    • 2015
  • High Efficiency Video Coding (HEVC) is the most recent video codec standard of the ITU-T Video Coding Experts Group and the ISO/IEC Moving Picture Experts Group. The main goal of this newly introduced standard is for catering to high-resolution video in low bandwidth environments with a higher compression ratio. This paper provides a performance comparison between HEVC and H.264/AVC video compression standards in terms of objective quality, delay, and complexity in the broadcasting environment. The experimental investigation was carried out using six test sequences in the random access configuration of the HEVC test model (HM), the HEVC reference software. This was also carried out in similar configuration settings of the Joint Scalable Video Module (JSVM), the official scalable H.264/AVC reference implementation, running on a single layer mode. According to the results obtained, the HM achieves more than double the compression ratio compared to that of JSVM and delivers the same video quality at half the bitrate. Yet, the HM encodes two times slower (at most) than JSVM. Hence, it can be concluded that the application scenarios of HM and JSVM should be judiciously selected considering the availability of system resources. For instance, HM is not suitable for low delay applications, but it can be used effectively in low bandwidth environments.

First Report of Tomato Spotted Wilt Virus on Iris domestica in South Korea (약용작물 범부채에 발생한 Tomato Spotted Wilt Virus 국내 첫 보고)

  • Chung, Bong Nam;Yoon, Ju-Yeon;Cho, In-Sook
    • Research in Plant Disease
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    • v.27 no.1
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    • pp.32-37
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    • 2021
  • In May 2020, necrosis and necrotic ring patterns were observed on leaves of three of 140 Iris domestica plants in a demonstration garden in Wanju, Jeollabuk-do. Three symptomatic plants were found to be infected by tomato spotted wilt virus (TSWV). To analyze the whole genomic sequence of one TSWV isolate, 'Blackberry lily-kr1', L, M, and S genome segments were sequenced and analyzed by comparison of nucleotide sequences of the three segments with corresponding sequences of other TSWV isolates. 'Blackberry lily-kr1' isolate was most closely related to 'JJ' isolate (MF159046) or 'HJ' isolate (LC273305) in the L segment, and to 'JJ' isolate (MF159058 and KY021439) in the M and S segments, respectively. Phylogenetic analysis by Maximum likelihood method using MEGA X program with 'Blackberry lily-kr1' isolate showed high relationship with 'JJ' pepper isolate or 'HJ' Humulus japonicas isolate in the all three segment. Necrosis and double ring patterns on leaves were formed in the glasshouse after inoculation of healthy I. domestica plants with sap of 'Blackberry lily-kr1'-infected Nicotiana rustica plants. This result suggests that I. domestica plants showing necrotic ring patterns in the open field are caused by TSWV infection. This is the first report of TSWV infection of I. domestica in Korea.

A Perceptual Rate Control Algorithm with S-JND Model for HEVC Encoder (S-JND 모델을 사용한 주관적인 율 제어 알고리즘 기반의 HEVC 부호화 방법)

  • Kim, JaeRyun;Ahn, Yong-Jo;Lim, Woong;Sim, Donggyu
    • Journal of Broadcast Engineering
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    • v.21 no.6
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    • pp.929-943
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    • 2016
  • This paper proposes the rate control algorithm based on the S-JND (Saliency-Just Noticeable Difference) model for considering perceptual visual quality. The proposed rate control algorithm employs the S-JND model to simultaneously reflect human visual sensitivity and human visual attention for considering characteristics of human visual system. During allocating bits for CTU (Coding Tree Unit) level in a rate control, the bit allocation model calculates the S-JND threshold of each CTU in a picture. The threshold of each CTU is used for adaptively allocating a proper number of bits; thus, the proposed bit allocation model can improve perceptual visual quality. For performance evaluation of the proposed algorithm, the proposed algorithm was implemented on HM 16.9 and tested for sequences in Class B and Class C under the CTC (Common Test Condition) RA (Random Access), Low-delay B and Low-delay P case. Experimental results show that the proposed method reduces the bit-rate of 2.3%, and improves BD-PSNR of 0.07dB and bit-rate accuracy of 0.06% on average. We achieved MOS improvement of 0.03 with the proposed method, compared with the conventional method based on DSCQS (Double Stimulus Continuous Quality Scale).

A GNSS Code Tracking Scheme Based in Slope Difference of Correlation Outputs (상관 함수의 기울기 차에 기반한 GNSS의 부호 추적 기법)

  • Yoo, Seung-Soo;Yoo, Seung-Hwan;Chong, Da-Hae;Ahn, Sang-Ho;Yoon, Seok-Ho;Kim, Sun-Yong
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.33 no.6C
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    • pp.505-511
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    • 2008
  • The global navigation satellite system (GNSS) is using a direct sequence/spread spectrum (DS/SS) modulation. In order to recover the information data, the DS/SS system first performs a two-step synchronization process: acquisition and tracking. The acquisition process adjusts the phase difference between the received and locally generated acquisition sequences within ${\pm}T_c/2$ or less, where $T_c$ is the chip period. The tracking process performs fine synchronization. In this paper, we focus on the tracking issue. The single delta delay locked loop($\Delta$-DLL) is the optimal tracking scheme for a GNSS in the absence of multipath signals, where $\Delta$ means the spacing between the early and late correlation time offset. In the multipath environments, however, the $\Delta$-DLL suffers from huge estimation bias(denoted by $\beta$) caused by distorted correlation values. Although some modified schemes such as a $\Delta$-DLL with a narrow $\Delta$ and a double delta DLL (${\Delta}^{(2)}$-DLL) were proposed to reduce the estimation bias, they cannot remove the estimation bias completely and need more accurate acquisition process. This paper proposes a novel tracking scheme that can dramatically reduce the estimation bias, using the maximum slope change among the correlation outputs.