• Title/Summary/Keyword: Hybrid sequencing

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Chemical Synthesis and Cloning of Panax ginseng Peptide Gene

  • Zhang, Hong-Ying;Chen, Dong-Song;Zhang, Jin
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.65-67
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    • 1990
  • The sequence of ginseng peptide gene was designed and synthesized by the solid phase plasmid pUC19. Escherichia coli JM101 cells were transformed with above hybrid plasmids. Ampicillin resistant transformants were screened and identified by in situ colony hybridization and Southern blot techinques. Finally the gene sequencing was done by the Sanger dideoxy method using primer extension.

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Chemical Synthesis and Cloning of Panax ginseng Peptide Gene (인삼펩티드 유전자의 합성 및 클로닝)

  • Zhang, Hong-Ying;Chen, Dong-Song;Zhang, Jin
    • Journal of Ginseng Research
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    • v.14 no.2
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    • pp.207-209
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    • 1990
  • The sequence of ginseng peptide gene was designed and synthesized by the solid phase phosphoramidiate method. Synthetic segments were isolated, pllrified and joined to the plasmid pUC19. E.icherichiu coli JM101 cells were transformed with above hybrid plasmids. AmpiciIBin resistant transformants were screened and identified by in situ colony hybridization and Southern blot techniques. Finally the gene sequencing was done by the Sanger dideoxy method using primer extension.

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Sequencing Strategy for Autonomous Mobile Robots in Real Environments (이동로봇 자율주행을 위한 행위모듈의 실행순서 조정기법)

  • 송인섭;박정민;오상록;조영조;박귀태
    • Journal of Institute of Control, Robotics and Systems
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    • v.5 no.3
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    • pp.297-305
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    • 1999
  • Autonomous mobile robots are required to achieve multiple goals while responding quickly to the dynamic environments. An appropriate robot control architecture, which clearly and systematically defines the relationship among the inputs, the processing functions and the outputs, thus needs to be embedded in the robot controller. This paper proposes a kind of hybrid control architecture which combines the key features of the two well-known robot control architectures; hierarchical and behavioral- based. The overall control architecture consists of three layers, i.e. the highest planner, the middle plan executor, and the lowest monitor and behavior-based controller. In the planned situation, only one behavior module is chosen by the logical coordinator in the plan executor according to the way point bin. In the exceptional situation, the central controller in the plan executor issues an additional control command to reach the planned way point. Several simulations and experiments with autonomous mobile robot show that the proposed architecture enables the robot controller to achieve the multiple sequential goals even in dynamic and uncertain environments.

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Amino Acid Sequence Homology of Hybrid Poplar O-methyltransferuse Involved in Lignin Biosynthesis

  • Park, Young-Goo;Sul, Ill-Whan;Shin, Dong-Ill;Park, Jang-Won;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.3 no.3
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    • pp.131-134
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    • 2001
  • In $\lambda$-Zap II vector system, a cDNA library was constructed for the developing secondary xylem mRNA from hybrid poplar, Populus nigra x maximowiczii. A cDNA clone of 1.5 kb in size, pOMTB1.4 encoding a lignin-bispecific O-methyltransferase was screened by plaque hybridization using a probe of 540 bp cDNA amplified by polymerase chain reaction from the cDNA library and identified by nucleotide sequencing. Its nucleotide sequence contains one open reading frame of 366 amino acids. The deduced amino acid sequence in comparison with that of Populus tremuloides showed the differences of 9 amino acids and revealed 85-99% homology among alfalfa, poplar and aspen.

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Interaction between HIV-1 Nef and LyF-1, the T Cell Specific Transcription Factor (T 세포 특이적 전사인자인 LyF-1과 HIV-1 Nef의 상호 작용)

  • Lee, Mi-Seon;Lee, Kyoung-Hoa;Kim, Jung-Woo
    • The Journal of Korean Society of Virology
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    • v.30 no.3
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    • pp.211-217
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    • 2000
  • Nef is a lentiviral protein involved in pathogenesis of AIDS, but its molecular mechanism of action remains incompletely understood. Here we report the isolation of the interacting protein with the HIV-1 Nef, using the yeast two hybrid system for expression cloning. One of the positive colonies was selected as the final candidate for the interacting protein gene. The nucleotide sequencing revealed that this interacting protein is Human Ikaros/LyF-1. This protein interacted with the C-terminal region of Nef specifically in yeast system, not with the N-terminal region. This interaction was also confirmed by in vitro binding assay.

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Performance Evaluation of Hybrid SBR Aerobic Digestion combined with Ultrasonication by using a Mathematical Model (초음파 결합형 SBR 호기성 소화 모델과 영향 예측)

  • Kim, Sung Hong;Lee, Dong Woo;Kim, Dong Han
    • Journal of Korean Society of Water and Wastewater
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    • v.26 no.6
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    • pp.897-905
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    • 2012
  • Based on the activated sludge model, a simple aerobic digestion model which represents the aerobic sludge digestion by sequencing batch reactor(SBR) equipped with ultrasonicator was composed and performed in this study. The results are as follows. Aerobic digestion efficiency can be increased by adopting ultrasonic pretreatment. For the 5 days of SRT, 24 % of particulate component is predicted to be removed by ultrasonic pretreatment and aerobic digestion. This is 7 %p higher than that of conventional aerobic digestion. A Hybrid SBR aerobic digestion combined with ultrasonication shows higher digestion efficiency than aerobic digestion and ultrasonic pretreatment system. In case of this hybrid system, the digestion efficiency was predicted up to 49 % when the ultrasonication was performed every 2 hours, repeatedly. However, excessive treatment like every hours of ultrasonication in an aerobic digestion process results in adverse effect on TCOD removal because biomass disintegrated completely and the solubilized COD can not be used for the biomass synthesis again.

Molecular Biological Species Identification of Imported Groupers(Epinephelus moara ♀×E. lanceolatus ♂) (수입산 바리과(Family Serranidae) 잡종 어류(Epinephelus moara ♀×E. lanceolatus ♂)의 분자생물학적 판별)

  • Kim, Yong Hwi;Park, Jong Yeon;Kim, Jae Hoon;Bang, In-Chul
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.4
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    • pp.566-571
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    • 2020
  • To classify a presumed hybrid of imported grouper species acquired from the National Fishery Products Quality Management Service, maternal and paternal lines were identified based on partial sequencing of mitochondrial cytochrome c oxidase 1 (co1) and nuclear recombination activation gene 1 (rag1) genes. The matrilineal species was identified as Epinephleus moara by a partial (760 bp) co1 sequence. Ambiguous sequences with base pairs belonging to E. moara or E. lanceolatus were found in a total of 15 different base pairs in the partial 1,159 bp of the rag1 gene, and the patrilineal species was found to be E. lanceolatus. Therefore, all of the groupers examined in the study were identified to be hybrids of E. moara and E. lanceolatus. In addition, a fast and convenient method using random amplification of polymorphic DNA (RAPD) was established for hybrid discrimination. Hybrids between E. moara ♀ and E. lanceolatus ♂ were identified through specific bands of 387 bp and 433 bp in PRIMER 6.

A new record of Ardisia×walkeri, a hybrid of A. japonica and A. pusilla, (Primulaceae) from Jeju Island, Korea

  • Goro Kokubugata;Satoshi Kakishima;Chan-ho Park;Takuro Ito;Atsushi Abe;Chikako Ishii;Gwan-Pil Song
    • Journal of Species Research
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    • v.12 no.3
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    • pp.258-265
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    • 2023
  • We conducted phylogenetic analyses using multiplexed inter-simple sequence repeat genotyping by sequencing and compared chloroplast DNA sequences among Ardisia japonica, A. pusilla, and morphologically intermediate plants found on Jeju Island, Korea. Our network analysis demonstrated that the intermediate plants were genetically positioned between A. japonica and A. pusilla. Our comparison of the intergenic spacer between the psbA and trnH genes in chloroplast DNA indicated that four nucleotide substitutions separate A. japonica and A. pusilla, whereas the intermediate plants exhibited the A. japonica haplotype. Our results suggest that the intermediate plants on Jeju Island represent a natural hybrid of A. japonica, as the maternal species, and A. pusilla, and that they are attributable to Ardisia×walkeri. This record constitutes the first documented occurrence of the hybrid taxon in Korea.

Derivation of Embryonic Germ Cells from Post Migratory Primordial Germ Cells, and Methylation Analysis of Their Imprinted Genes by Bisulfite Genomic Sequencing

  • Shim, Sang Woo;Han, Dong Wook;Yang, Ji Hoon;Lee, Bo Yeon;Kim, Seung Bo;Shim, Hosup;Lee, Hoon Taek
    • Molecules and Cells
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    • v.25 no.3
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    • pp.358-367
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    • 2008
  • The embryonic germ cell (EGCs) of mice is a kind of pluripotent stem cell that can be generated from pre- and post-migratory primordial germ cells (PGCs). Most previous studies on DNA methylation of EGCs were restricted to 12.5 days post coitum (dpc). This study was designed to establish and characterize murine EGC lines from migrated PGCs as late as 13.5 dpc and to estimate the degrees of methylation of their imprinted genes as well as of the non-imprinted locus, Oct4, using an accurate and quantitative method of measurement. We established five independent EGC lines from post migratory PGCs of 11.5-13.5 dpc from C57BL/6 ${\times}$ DBA/2 F1 hybrid mouse fetuses. All the EGCs exhibited the typical features of pluripotent cells including hypomethylation of the Oct4 regulatory region. We examined the methylation status of three imprinted genes; Igf2, Igf2r and H19 in the five EGC lines using bisulfite genomic sequencing analysis. Igf2r was almost unmethylated in all the EGC lines irrespective of the their sex and stage of isolation; Igf2 and H19 were more methylated than Igf2r, especially in male EGCs. Moreover, EGCs derived at 13.5 dpc exhibited higher levels of DNA methylation than those from earlier stages. These results suggest that in vitro derived EGCs acquire different epigenotypes from their parental in vivo migratory PGCs, and that sex-specific de novo methylation occurs in the Igf2 and H19 genes of EGCs.

Survey of the Applications of NGS to Whole-Genome Sequencing and Expression Profiling

  • Lim, Jong-Sung;Choi, Beom-Soon;Lee, Jeong-Soo;Shin, Chan-Seok;Yang, Tae-Jin;Rhee, Jae-Sung;Lee, Jae-Seong;Choi, Ik-Young
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.1-8
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    • 2012
  • Recently, the technologies of DNA sequence variation and gene expression profiling have been used widely as approaches in the expertise of genome biology and genetics. The application to genome study has been particularly developed with the introduction of the nextgeneration DNA sequencer (NGS) Roche/454 and Illumina/ Solexa systems, along with bioinformation analysis technologies of whole-genome $de$ $novo$ assembly, expression profiling, DNA variation discovery, and genotyping. Both massive whole-genome shotgun paired-end sequencing and mate paired-end sequencing data are important steps for constructing $de$ $novo$ assembly of novel genome sequencing data. It is necessary to have DNA sequence information from a multiplatform NGS with at least $2{\times}$ and $30{\times}$ depth sequence of genome coverage using Roche/454 and Illumina/Solexa, respectively, for effective an way of de novo assembly. Massive shortlength reading data from the Illumina/Solexa system is enough to discover DNA variation, resulting in reducing the cost of DNA sequencing. Whole-genome expression profile data are useful to approach genome system biology with quantification of expressed RNAs from a wholegenome transcriptome, depending on the tissue samples. The hybrid mRNA sequences from Rohce/454 and Illumina/Solexa are more powerful to find novel genes through $de$ $novo$ assembly in any whole-genome sequenced species. The $20{\times}$ and $50{\times}$ coverage of the estimated transcriptome sequences using Roche/454 and Illumina/Solexa, respectively, is effective to create novel expressed reference sequences. However, only an average $30{\times}$ coverage of a transcriptome with short read sequences of Illumina/Solexa is enough to check expression quantification, compared to the reference expressed sequence tag sequence.