• Title/Summary/Keyword: Sequence Selection

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Isolation and Characterization of Exopolysaccharide-Producing Bacteria from Korean Fermented Vegetables (전통 침채류 유래 다당 생산균의 분리 특성)

  • Kwon, Tae-Yeon;Shim, Sang-Min;Heo, Min-Young;An, Doo-Hyun;Shin, Kwang-Soon;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.191-195
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    • 2007
  • Four bacteria producing viscous exopolysaccharides (EPSs) were isolated from Korean fermented vegetables (Cucumber kimchi, Young radish kimchi, Green onion kimchi) using a selection medium intended for isolating bacteria with tannin-degrading activity. They were identified phylogenetically by 16S rDNA sequence analysis and found to be very close to Enterobacter cowan ii, Escherichia senegalensis, Enterobacter asburiae, and Enterobacter ludwigii. Strain CK31, the most efficient EPS-producer, produced a heteropolysaccharide with an approximate molecular weight of 420 kDa. The neutral sugar fraction of the EPS was composed of rhamnose, fucose, arabinose, mannose, galactose, and glucose.

Probiotic Properties of Lactobacillus salivarius CPM-7 Isolated from Chicken Feces. (계분으로부터 Lactobacillus salivarius의 분리 및 생균제적 특성)

  • Lim, Soo-Jin;Jang, Sung-Sik;Kang, Dae-Kyung
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.98-103
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    • 2007
  • To isolate probiotic lactic acid bacteria for animal, we have screened the microorganisms from chicken feces, by random selection and agar well diffusion assay. Among them, CPM-7 strain showing superior inhibitory activity against Escherichia coli was selected. By examining carbohydrates utilization, morphologic property and 16S rRNA gene sequence, CPM-7 strain was identified as Lactobacillus salivarius, then named L. salivarius CPM-7. L. salivarius CPM-7 produced thirteen enzymes in the test using API ZYM kit, and showed resistance to low pH and bile salts. It survived at pH 2 for 30 min. and pH 3 for 6 hr. And, it was able to grow in MRS medium containing 0.2% (w/v) bile salts. L. salivarius CPM-7 adhered to the jejunal epithelium cells of pig. Both the supernatant of L. salivarius CPM-7 and the its neutralized one showed high inhibitory activity against E. coli K88.

Triclosan Resistance in a Bacterial Fish Pathogen, Aeromonas salmonicida subsp. salmonicida, is Mediated by an Enoyl Reductase, FabV

  • Khan, Raees;Lee, Myung Hwan;Joo, Haejin;Jung, Yong-Hoon;Ahmad, Shabir;Choi, Jinhee;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.511-520
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    • 2015
  • Triclosan, the widely used biocide, specifically targets enoyl-acyl carrier protein reductase (ENR) in the bacterial fatty acid synthesis system. Although the fish pathogen Aeromonas salmonicida subsp. salmonicida exhibits triclosan resistance, the nature of this resistance has not been elucidated. Here, we aimed to characterize the triclosan resistance of A. salmonicida subsp. salmonicida causing furunculosis. The fosmid library of triclosan-resistant A. salmonicida subsp. salmonicida was constructed to select a fosmid clone showing triclosan resistance. With the fosmid clone showing triclosan resistance, a subsequent secondary library search resulted in the selection of subclone pTSR-1. DNA sequence analysis of pTSR-1 revealed the presence of a chromosomal-borne fabV-encoding ENR homolog. The ENR of A. salmonicida (FabVas) exhibited significant homology with previously known FabV, including the catalytic domain YX(8)K. fabVas introduction into E. coli dramatically increased its resistance to triclosan. Heterologous expression of FabVas might functionally replace the triclosan-sensitive FabI in vivo to confer E. coli with triclosan resistance. A genome-wide search for fabVas homologs revealed the presence of an additional fabV gene (fabVas2) paralog in A. salmonicida strains and the fabVas orthologs from other gram-negative fish pathogens. Both of the potential FabV ENRs expressed similarly with or without triclosan supplement. This is the first report about the presence of two potential FabV ENRs in a single pathogenic bacterium. Our result suggests that triclosan-resistant ENRs are widely distributed in various bacteria in nature, and the wide use of this biocide can spread these triclosan-tolerant ENRs among fish pathogens and other pathogenic bacteria.

A Novel Integrative Expression Vector for Sulfolobus Species

  • Choi, Kyoung-Hwa;Hwang, Sungmin;Yoon, Naeun;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1503-1509
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    • 2014
  • With the purpose of facilitating the process of stable strain generation, a shuttle vector for integration of genes via a double recombination event into two ectopic sites on the Sulfolobus acidocaldarius chromosome was constructed. The novel chromosomal integration and expression vector pINEX contains a pyrE gene from S. solfataricus P2 ($pyrE_{sso}$) as an auxotrophic selection marker, a multiple cloning site with histidine tag, the internal sequences of malE and malG for homologous recombination, and the entire region of pGEM-T vector, except for the multiple cloning region, for propagation in E. coli. For stable expression of the target gene, an ${\alpha}$-glucosidase-producing strain of S. acidocaldarius was generated employing this vector. The malA gene (saci_1160) encoding an ${\alpha}$-glucosidase from S. acidocaldarius fused with the glutamate dehydrogenase ($gdhA_{saci}$) promoter and leader sequence was ligated to pINEX to generate pINEX_malA. Using the "pop-in" and "pop-out" method, the malA gene was inserted into the genome of MR31 and correct insertion was verified by colony PCR and sequencing. This strain was grown in YT medium without uracil and purified by His-tag affinity chromatography. The ${\alpha}$-glucosidase activity was confirmed by the hydrolysis of $pNP{\alpha}G$. The pINEX vector should be applicable in delineating gene functions in this organism.

Development of a Genome-Wide Random Mutagenesis System Using Proofreading-Deficient DNA Polymerase ${\delta}$ in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Oh Cheol;Kim, Sang-Yoon;Hwang, Dong Hyeon;Oh, Doo-Byoung;Kang, Hyun Ah;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.304-312
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    • 2013
  • The thermotolerant methylotrophic yeast Hansenula polymorpha is attracting interest as a potential strain for the production of recombinant proteins and biofuels. However, only limited numbers of genome engineering tools are currently available for H. polymorpha. In the present study, we identified the HpPOL3 gene encoding the catalytic subunit of DNA polymerase ${\delta}$ of H. polymorpha and mutated the sequence encoding conserved amino acid residues that are important for its proofreading 3'${\rightarrow}$5' exonuclease activity. The resulting $HpPOL3^*$ gene encoding the error-prone proofreading-deficient DNA polymerase ${\delta}$ was cloned under a methanol oxidase promoter to construct the mutator plasmid pHIF8, which also contains additional elements for site-specific chromosomal integration, selection, and excision. In a H. polymorpha mutator strain chromosomally integrated with pHIF8, a $URA3^-$ mutant resistant to 5-fluoroorotic acid was generated at a 50-fold higher frequency than in the wild-type strain, due to the dominant negative expression of $HpPOL3^*$. Moreover, after obtaining the desired mutant, the mutator allele was readily removed from the chromosome by homologous recombination to avoid the uncontrolled accumulation of additional mutations. Our mutator system, which depends on the accumulation of random mutations that are incorporated during DNA replication, will be useful to generate strains with mutant phenotypes, especially those related to unknown or multiple genes on the chromosome.

Improved changed region detection and motion estimation for object-oriented coding (객체기반 부호화에서의 개선된 움직임 영역 추출 및 추정 기법)

  • 정의윤;박영식;송근원;한규필;하영호
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.22 no.9
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    • pp.2043-2052
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    • 1997
  • The object-oriented coding technique which is one of the coding methods in very low bit rate environment is suitable for videophone image sequence. The selection of source model affect image analysis. In this paper, an image analysis method for the object-oriented coding is presented. The process is composed of changed region detection andmotion estimateion. First, we use the standard deviation of frame difference as thrreshold to extract themoving area. If thesum of gray values in mask is greater than the threshold, the center pixel of the mask is regarded as moving region. After moving is detected in changed region by edge operator, observation point is determined from moving region. The motion is estimated by 6-parameter mapping method with determined observation point. The experimantal resutls show that the proposed method can significantly improve the image quality.

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Putative Secondary Structure of Human Hepatitis B Viral X mRNA

  • Kim, Ha-Dong;Choi, Yoon-Chul;Lee, Bum-Yong;Junn, Eun-Sung;Ahn, Jeong-Keun;Kang, Chang-Won;Park, In-Won
    • BMB Reports
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    • v.28 no.6
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    • pp.509-514
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    • 1995
  • A putative secondary structure of the mRNA for the human hepatitis B virus (HBV) X gene is proposed based on not only chemical and enzymatic determination of its single- and double-stranded regions but also selection by the computer program MFOLD for energy minimum conformation under the constraints that the experimentally determined nucleotides were forced or prohibited to base pair. An RNA of 536 nucleotides including the 461-nucleotide HBV X mRNA sequence was synthesized in vitro by the phage T7 RNA polymerase transcription. The thermally renatured transcripts were subjected to chemical modifications with dimethylsulfate and kethoxal and enzymatic hydrolysis with single strand-specific RNase T1 and double strand-specific RNase V1, separately. The sites of modification and cleavage were detected by reverse transcriptase extension of 4 different primers. Many nucleotides could be assigned with high confidence, twenty in double-stranded and thirty-seven in Single-stranded regions. These nucleotides were forced and prohibited, respectively, to base pair in running the recursive RNA folding program MFOLD. The results suggest that 6 different regions (5 within X mRNA) of 14~23 nucleotides are Single-stranded. This putative structure provides a good working model and suggests potential target sites for antisense and ribozyme inhibitors and hybridization probes for the HBV X mRNA.

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Analysis on Flow Control Method for Simultaneous Fuel Filling of the Korea Space Launch Vehicle-II (한국형발사체 연료 동시충전을 위한 유량제어 방식에 대한 고찰)

  • Yeo, Inseok;Lee, Jaejun;An, Jaechel;Kang, Sunil
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2017.05a
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    • pp.5-13
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    • 2017
  • To lunch the Korea Space Launch Vehicle-II(KSLV-II), the second launch complex will be constructed on the Naro Space Center and Kerosene Filling System (KFS) will be also installed newly. KFS of KSLV-II launch complex system is being designed based on Naro Launch Complex. But this must supply fuel to fuel tanks of the vehicle with only a supply pump because KSLV-II is a 3-stage launch vehicle unlike Naro Launch Vehicle or Test Launch Vehicle (TLV). A sudden rise of pump output pressure is recognized during fuel filling scenario selection process. This occurs because return flow can not actively deal with a lot of flow change using flow control method of orifice type. To solve this problem, it is verified that fuel can be stably supplied by installation of accumulator and an appropriate adjustment of filling mode change sequence through flow analysis of various cases.

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Design and Construction of the Burj Dubai Concrete Building Project (버즈 두바이 콘크리트 건물의 설계와 시공)

  • Abdelrazaq, Ahmad
    • Magazine of the Korea Concrete Institute
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    • v.20 no.6
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    • pp.28-35
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    • 2008
  • The Burj Dubai Project will be the tallest structure ever built by man; when completed the tower will be more than 700 meter tall and more than 160 floors. While the early integration of aerodynamic shaping and wind engineering considerations played a major role in the architectural massing and design of this multi-use/residential tower, where mitigating and taming the dynamic wind effects was one of the most important design criteria, the material selection for the structural systems of the tower was also a major consideration and required detailed evaluation of the material technologies and skilled labor available in the market at the time Concrete was selected for its strength, stiffness, damping, redundancy, moldability, free fireproofing, speed of construction, and cost effectiveness. In addition, the design challenges of using concrete for the design of the structural system components will be addressed. The focus on this paper will also be on the early planning of the concrete works of the Burj Dubai Project.

Modeling and Direct Power Control Method of Vienna Rectifiers Using the Sliding Mode Control Approach

  • Ma, Hui;Xie, Yunxiang;Sun, Biaoguang;Mo, Lingjun
    • Journal of Power Electronics
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    • v.15 no.1
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    • pp.190-201
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    • 2015
  • This paper uses the switching function approach to present a simple state model of the Vienna-type rectifier. The approach introduces the relationship between the DC-link neutral point voltage and the AC side phase currents. A novel direct power control (DPC) strategy, which is based on the sliding mode control (SMC) for Vienna I rectifiers, is developed using the proposed power model in the stationary ${\alpha}-{\beta}$ reference frames. The SMC-based DPC methodology directly regulates instantaneous active and reactive powers without transforming to a synchronous rotating coordinate reference frame or a tracking phase angle of grid voltage. Moreover, the required rectifier control voltages are directly calculated by utilizing the non-linear SMC scheme. Theoretically, active and reactive power flows are controlled without ripple or cross coupling. Furthermore, the fixed-switching frequency is obtained by employing the simplified space vector modulation (SVM). SVM solves the complicated designing problem of the AC harmonic filter. The simplified SVM is based on the simplification of the space vector diagram of a three-level converter into that of a two-level converter. The dwelling time calculation and switching sequence selection are easily implemented like those in the conventional two-level rectifier. Replacing the current control loops with power control loops simplifies the system design and enhances the transient performance. The simulation models in MATLAB/Simulink and the digital signal processor-controlled 1.5 kW Vienna-type rectifier are used to verify the fast responses and robustness of the proposed control scheme.