• Title/Summary/Keyword: conserved

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A Myostain-like Gene Expressed Highly in the Muscle Tissue of Chinese mitten crab, Eriocheir sinensis

  • Kim, Kyoung-Sun;Jeon, Jeong-Min;Kim, Hyun-Woo
    • Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.185-193
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    • 2009
  • A complete cDNA, which encodes for a myostatin-like protein (Es-MSTN), was isolated from the Chinese mitten crab, Eriocheir sinensis. Es-MSTN was composed of 2,397 nucleotides and the open reading frame (ORF) specified a protein containing 468 amino acids. Es-MSTN exhibited 32% amino acid sequence identity and 52% similarity to human myostatin. Multiple sequence alignment analysis indicated that Es-MSTN possessed the conserved proteolytic cleavage site (RXXR) for maturation of the protein and nine cysteine residues for disulfide bridges. Besides the conserved structural features, Es-MSTN also exhibits its unique characters; a longer N-terminal domain which is involved in protein folding and latent form of myostatin and absence of the cleavage site for BMP-1/tolloid family of metalloproteinase to activate mature myostatin. Phylogenetic analysis suggests that Es-MSTN showed the closely related to both vertebrate myostatin and GDF11. Es-MSTN is expressed highly in the claw muscle, leg muscle, thoracic muscle and heart, and moderately in the hindgut suggesting that Es-MSTN may play important roles in the muscle tissues. As homolog of mammalian myostatin and GDF11, Es-MSTN may be involved in development of muscular tissue and further study will help to produce high-quality seafood.

Rapid and Efficient Isolation of Genes for Biosynthesis of Peptide Antibiotics from Gram-positive Bacterial Strains

  • Lee, Soon-Youl;Rhee, Sang-Ki;Kim, Chul-Ho;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.310-317
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    • 1998
  • Peptide synthetases are large multifunctional enzyme complexes that catalyze the nonribosomal synthesis of a structurally diverse family of peptide antibiotics. These enzymes are composed of functionally independent domains with independent enzymatic activities. Their specific linkage order of domains forms the protein template that defines the sequence of the incorporated amino acids. Within each domain, several motifs of highly conserved sequences have been identified from the sequence alignment of the various peptide synthetases [30]. Taking advantage of the conserved nucleotide sequence of Core 1 and Core 2, we designed PCR primers to amplify the peptide synthetase genes from three different gram-positive bacterial strains. Nucleotide sequence analysis of the amplified PCR products from those three strains showed significant homology to various peptide synthetase genes, suggesting that the PCR products are parts of peptide synthetase genes. Therefore, this rapid and efficient PCR technique can be used for the isolation of peptide synthetase genes from various strains.

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A Cloning of Novel Esterase from a Metagenomic Library

  • Yoon, Sang-Young;Kim, Seung-Bum;Ryu, Yeon-Woo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.243-246
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    • 2005
  • A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

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DNA Chip을 이용한 Transcriptional Activation Mechanism 분석

  • 김영준
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.45-60
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    • 2001
  • . Mediator of transcriptional regulation is the evolutionary conserved coactivator complex that plays He central role in the integration and recruitment of diverse regulatory signals and transcription machinery to certain promoters. In yeast, each Mediator subunit is required for transcriptional regulation of a distinct group of genes. In order to decipher the mechanistic roles of Mediator proteins in regulating developmental specific gene expression, we isolated, and analyzed a multiprotein complex containing Drosophila Mediate. homologs (dMediato.). dMediato. interacts with several sequence-sperific transcription factors and basal transcription machinery, and is critical for activated transcription in response to diverse transcriptional activators. In order to elucidate the function of Mediator in metazoan development, we isolated mutants of a conserved Mediate. subunit, Drosophila Med6 (dMed6). dMed6 null homozygotes failed to pupate and died in the third larval instar. Larval mitotic cells and most imaginal discs showed severe defects in proliferation, but no apparent morphological defect was observed in other larval tissues. Clonal analysis of dMed6 mutant cells revealed that dMed6 is essential for cell viability and proliferation of most adult cell types. Drosophila cDNA microarray, quantitative RT-PCR, and in situ expression analyses of developmentally regulated genes in dMed6 mutants showed that transcriptional activation of a subset of genes involved in neuroblast proliferation in the larval brain were most affected. Our results suggest that dMed6 is required in most for transcriptional regulation of a subset of genes important for cell proliferation and metabolism.

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Neural Substrates of Fear Based on Animal and Human Studies (공포의 신경 기저 회로 : 동물과 인간 대상 연구를 중심으로)

  • Baek, Kwangyeol;Jeong, Jaeseung;Park, Min-Sun;Chae, Jeong-Ho
    • Korean Journal of Biological Psychiatry
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    • v.15 no.4
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    • pp.254-264
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    • 2008
  • Objectives : The neural substrate of fear is thought to be highly conserved among species including human. The purpose of this review was to address the neural substrates of fear based on recent findings obtained from animal and human studies. Methods : Recent studies on brain regions related to fear, particularly fear conditioning in rodents and humans, were extensively reviewed. Results : This paper suggests high consistency in anatomical structure and physiological mechanisms for fear perception, response, learning and modulation in animals and humans. Conclusions : Fear is manifested and modulated by well conserved neural circuits among species interconnected with the amygdala, such as the hippocampus and the ventromedial prefrontal cortex. Further research is required to incorporate findings from animal studies into a better understanding of neural circuitry of fear in human in a translational approach.

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Isolation, Restriction Mapping, and Promoter Sequence Analysis of an Isoperoxidase Gene from Korean-Radish, Raphanus sativus L.

  • Park, Jong-Hoon;Kim, Soung-Soo
    • BMB Reports
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    • v.29 no.1
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    • pp.52-57
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    • 1996
  • A specific DNA fragment from Korean radish (Raphanus sativus L.) was amplified by performing PCR with oligonucleotide primers which correspond to the highly conserved regions of plant peroxidases. The size of the PCR product was ca. 400 bp, as expected from the known plant peroxidase genes. Comparison of the nucleotide and deduced amino acid sequences of the PCR product to those of other plant peroxidase-encoding genes revealed that the amplified fragment corresponded to the highly conserved region I and III of plant peroxidases. By screening a genomic library of Korean radish using the amplified fragment as a probe, two positive clones, named prxK1 and prxK2, were isolated. Restriction mapping studies indicated that the 5.2 kb Sail fragment of the prxK1 clone and the 4.0 kb EcoRI fragment of the prxK2 clone encode separate isoperoxidase genes. Analyses of the promoter region of the prxK1 clone shows that putative CAAT box, CMT box, and TGA1b binding sequence (5' TGACGT) are present 718 bp upstream from the start codon.

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Drosophila as a model for unfolded protein response research

  • Ryoo, Hyung Don
    • BMB Reports
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    • v.48 no.8
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    • pp.445-453
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    • 2015
  • Endoplasmic Reticulum (ER) is an organelle where most secretory and membrane proteins are synthesized, folded, and undergo further maturation. As numerous conditions can perturb such ER function, eukaryotic cells are equipped with responsive signaling pathways, widely referred to as the Unfolded Protein Response (UPR). Chronic conditions of ER stress that cannot be fully resolved by UPR, or conditions that impair UPR signaling itself, are associated with many metabolic and degenerative diseases. In recent years, Drosophila has been actively employed to study such connections between UPR and disease. Notably, the UPR pathways are largely conserved between Drosophila and humans, and the mediating genes are essential for development in both organisms, indicating their requirement to resolve inherent stress. By now, many Drosophila mutations are known to impose stress in the ER, and a number of these appear similar to those that underlie human diseases. In addition, studies have employed the strategy of overexpressing human mutations in Drosophila tissues to perform genetic modifier screens. The fact that the basic UPR pathways are conserved, together with the availability of many human disease models in this organism, makes Drosophila a powerful tool for studying human disease mechanisms. [BMB Reports 2015; 48(8): 445-453]

Implicit/Explicit Finite Element Method for Euler Flows Inside the Optimum Nozzle (내/외재적 유한요소법을 이용한 최대추력노즐의 설계해석)

  • Yoon W. S.;Kho H.
    • Journal of computational fluids engineering
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    • v.2 no.1
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    • pp.66-72
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    • 1997
  • Optimum nozzle design exploiting the method of characteristic(M.O.C) has been in application as an efficient design methodology targeting a less weighted and short expansion nozzle. This paper treats the optimum nozzle design and the analysis of the inviscid compressible flow inside. Based on traditional Rao's method, the optimum nozzle design is coded with minor modifications for the identification of the control surface across which the mass flux should be conserved. Internal flow field is simulated numerically by M.O.C and implicit/explicit Taylor-Galerkin finite element method(F.E.M) with the aid of adaptive remeshing to capture the shock wave, hence improve the accuracy. Designed and calculated flow fields due to the separate analyses show that the mass flux predicted by optimum nozzle design with M.O.C is not conserved across the control surface and the sonic line should be located upstream of the nozzle throat. Rao's optimum nozzle design methodology exaggerates the momentum thrust and tends to overemphasize the engine performance loss.

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Mutational Analysis of Two Conserved Active Site Tyrosine Residues in Matrilysin

  • Jaeho Cha
    • Journal of Life Science
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    • v.9 no.2
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    • pp.44-48
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    • 1999
  • The ionization of tyrosine residue is known to be involved in the stabilization of transition-state in catalysis of astacin based upon the astacin-transition state analog structure. Two tyrosine residues, Tyr-216 and Tyr-219, are conserved in all MMPs related with astacin family, We replaced Tyr-216 and Tyr-219 into phenylalanine, respectively and the zinc binding properties, kinetic parameters, and pH dependence of each mutant are determined in order to examine the role of tyrosine residue in matrilysin catalysis. Both mutants contain two zinc atoms per mol of enzyme, indicating that either tyrosime does not affect the zinc binding property of the enzyme. Y216F and Y219F mutants are highly active and the kcat/Km values are only decreased 1.1-1.5-fold compared to the wild-type enzyme. The decrease in the activity of the mutants is essentially due to the increase in Km value. The pH dependencies of the kcat/Km values for both mutants are similar to the corresponding dependencies obtained with the wild type enzyme. The pKa values at the alkaline side of both mutants are not changed. These kinetic and pH dependence results indicate that the ionization of active site tyrosine residue of matrilysin is not reflected in the kinetics of peptide hydrolysin as catalyzed by astacin.

Analysis of Flows in the Combustor with Recirculating Flow Regime (재순환영역을 가지는 연소기내의 연소유동해석)

  • 신동신;허남건
    • Journal of the Korean Society of Propulsion Engineers
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    • v.1 no.2
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    • pp.22-31
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    • 1997
  • We developed a general purpose program for the analysis of flows in the combustor with recirculating flow regime and simulated the flows. The program uses non-staggered grids based on finite volume method and the primitive variables are cartesian velocities. The combustion model is irreversible one step reaction with infinite chemistry The Favre averaged governing equations are considered and the clipped gaussian distribution is considered as a probability density function of the conserved scalar. We calculated turbulent diffusion flame with recirculating flow regime. Simulation shows two recirculating regions like experimental results. Velocity, turbulent kinetic energy, temperature and concentration distribution in simulation agree well with experimental data.

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