• Title/Summary/Keyword: Construction Sequence Analysis

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Construction of Transformation Method for Streptomyces scabiei ATCC 49173 Producing Phytotoxin (식물독소를 생산하는 Streptomyces scabiei ATCC 49173의 형질전환법 구축)

  • Jang, Bo-Youn;Ha, Heon-Su;Choi, Sun-Uk
    • KSBB Journal
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    • v.25 no.2
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    • pp.167-172
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    • 2010
  • Streptomyces scabiei producing phytotoxin called thaxtomin, which cause scab disease on economically important crops such as potato. For molecular genetics study of S. scabiei an effective transformation method was established based on conjugal transfer from Escherichia coli ET12567 (pUZ8002) using a phiC31-derived integration vector, pSET152, containing oriT and attP fragments. The high frequency was obtained on MS medium containing 50 mM $MgCl_2$. In addition, the sequence and location of the chromosomal integration attB site of S. scabiei was identified for the first time in the strains producing thaxtomin by the southern blot analysis of exconjugants and the sequencing of plasmid containing DNA flanking the insertion sites from exconjugant chromosome. Similar to the case of Streptomyces species, a single phiC31 attB site of S. scabiei is present within an ORF encoding a pirin-homolog.

Construction of Stably Transformed Bm5 Cells by Using Autographa californica Nuclear Polyhedrosis Virus IE1 Gene

  • Cho, Eun-Sook;Jin, Byung-Rae;Sohn, Hung-Dae;Chol, Kwang-Ho;Kim, Soung-Ryul;Kang, Seok-Woo;Yun, Eun-Young;Kim, Sang-Hyun;Kim, Keun-Young
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.111-116
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    • 1998
  • To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

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Start and Stop Characteristics of Single-Rod Electro-Hydrostatic Actuator (전동기 일체형 편로드 유압액추에이터의 기동 및 정지특성해석)

  • Jung, Gyu-Hong
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.35 no.11
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    • pp.1483-1490
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    • 2011
  • Electro-hydrostatic actuators(EHAs), which are usually composed of a direct motor-driven hydraulic pump and a cylinder, have been widely adopted as aircraft actuation systems because of their benefits in terms of improved efficiency, weight savings and the fact that they use a standalone power source. Since the recent trend in construction vehicles has been focus on energy savings in their hydraulic systems, EHAs are expected to be potential substitutes for conventional power transmission, since they are capable of energy recovery as well as highly efficient pump control. In this paper, the start and stop characteristics of EHAs were investigated through cracking pressure analysis of the pilot-operated check valve(PCV), which enables the cylinder to standstill against an external load with no holding effort from the hydraulic pump. A mathematical model that includes the load dynamics and the EHA's internal hydraulic circuit was derived for simulation with the MATLAB Simulink package. This model verified the PCV's opening and closing sequence, which in turn affects the EHA's start and stop characteristics.

Modification of Unit-Segmenting Schemes for Division Problems Involving Fractional Quantities (단위분할 도식의 재구성을 통한 포함제 분수나눗셈 문제해결에 관한 연구)

  • Shin, Jae-Hong;Lee, Soo-Jin
    • School Mathematics
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    • v.14 no.2
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    • pp.191-212
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    • 2012
  • In the field of arithmetic in mathematics education, there has been lack of fine-grained investigations addressing the relationship between students' construction of division knowledge with fractional quantities and their whole number division knowledge. This study, through the analysis of part of collected data from a year-long teaching experiment, presents a possible constructive itinerary as to how a student could modify her unit-segmenting scheme to deal with various fraction measurement division situations: 1) unit-segmenting scheme with a remainder, 2) fractional unit-segmenting scheme. Thus, this study provides a clue for curing a fragmentary approach to teaching whole number division and fraction division and preventing students' fragmentary understanding of the same arithmetical operation in different number systems.

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Radiative transfer analysis for Amon-Ra instrument

  • Seong, Se-Hyun;Ryu, Dong-Ok;Lee, Jae-Min;Hong, Jin-Suk;Kim, Seong-Hui;Yoon, Jee-Yeon;Park, Won-Hyun;Lee, Han-Shin;Park, Jong-Soo;Yu, Ji-Woong;Kim, Sug-Whan
    • Bulletin of the Korean Space Science Society
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    • 2009.10a
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    • pp.28.4-29
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    • 2009
  • The 'Amon-Ra' instrument of the proposed 'EARTHSHINE' satellite is a dual (i.e. imaging and energy) channel instrument for monitoring the total solar irradiance (TSI) and the Earth's irradiance at around the L1 halo orbit. Earlier studies for this instrument include, but not limited to, design and construction of breadboard Amon-Ra imaging channel, stray light suppression and system performance computation using Integrated Ray Tracing (IRT) technique. The Amon-Ra instrument is required to produce 0.3% in uncertainty for both Sunlight and Earthlight measurement. In this study, we report accurate estimation of the output electric signal derived from the orbital variation of radiant exitance from the Sun and the Earth arriving at the aperture and detector plane of the Amon-Ra. For this, orbital irradiance are computed analytically first and then confirmed by simulation using Integrated Ray Tracing (IRT) model. Specially, the results show the arriving power at the bolometer detector surface is $1.24{\mu}W$ for the Sunlight and $1.28{\mu}W$ for the Earthlight, producing the output signal pulses of 34.31 mV and 35.47 mV respectively. These results demonstrate successfully that the arriving radiative power is well within the bolometer detector dynamic range and, therefore, the proposed detector can be used for the in-orbit measurement sequence. We discuss the computational details and implications as well as the simulation results.

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Identification and Functional Analysis of Vibrio vulnificus SmcR, a Novel Global Regulator

  • Lee, Jeojng-Hyun;Rhee, Jee-Eun;Park, U-Ryung;Ju, Hyun-Mok;Lee, Byung-Cheol;Kim, Tae-Sung;Jeong, Hye-Sook;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.325-334
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    • 2007
  • Recently, quorum sensing has been implicated as an important global regulator controlling the production of numerous virulence factors such as capsular polysaccharides in bacterial pathogens. The nucleotide and deduced amino acid sequences of smcR, a homolog of V. harveyi luxR identified from V. vulnificus ATCC29307, were analyzed. The amino acid sequence of SmcR from V. vulnificus was 72 to 92% similar to those of LuxR homologs from Vibrio spp. Functions of SmcR were assessed by the construction of an isogenic mutant, whose smcR gene was inactivated by allelic exchanges, and by evaluating its phenotype changes in vitro and in mice. The disruption of smcR resulted in a significant alteration in biofilm formation, in type of colony morphology, and in motility. When compared with the wild-type, the smcR mutant exhibited reduced survival under adverse conditions, such as acidic pH and hyperosmotic stress. The smcR mutant exhibited decreased cytotoxic activity toward INT 407 cells in vitro. Furthermore, the intraperitoneal $LD_{50}$ of the smcR mutant was approximately $10^2$ times higher than that of parental wild-type. Therefore, it appears that SmcR is a novel global regulator, controlling numerous genes contributing to the pathogenesis as well as survival of V. vulnificus.

Investigation on Support Mechanism of Geogrid-Encased Stone Columns in Soft Ground (연약지반에 시공되는 지오그리드 감쌈 스톤컬럼의 하중 지지 메카니즘에 관한 연구)

  • Yoo, Chung-Sik;Kim, Sun-Bin
    • Journal of the Korean Geotechnical Society
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    • v.24 no.12
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    • pp.93-101
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    • 2008
  • This paper presents the results of numerical investigation on support mechanism of geogrid-encased stone columns for use in soft ground improvement. A number of cases were analyzed using a 3D stress-pore pressure coupled model that can effectively model construction sequence and drainage as well as reinforcing effects of geogrid-encased stone columns. The results indicated that the geogrid encasement provides additional confinement effect that reduces vertical stress in the soft ground, thus resulting in less excess pore water pressures and associated settlement. Also revealed was that such a confinement effect depends on encasement length and stiffness of geogrid. It is also shown that there exist critical encasement length and stiffness of geogrid for a given condition.

Full-Length Enriched cDNA Library Construction from Tissues Related to Energy Metabolism in Pigs

  • Lee, Kyung-Tai;Byun, Mi-Jeong;Lim, Dajeong;Kang, Kyung-Soo;Kim, Nam-Soon;Oh, Jung-Hwa;Chung, Chung-Soo;Park, Hae-Suk;Shin, Younhee;Kim, Tae-Hun
    • Molecules and Cells
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    • v.28 no.6
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    • pp.529-536
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    • 2009
  • Genome sequencing of the pig is being accelerated because of its importance as an evolutionary and biomedical model animal as well as a major livestock animal. However, information on expressed porcine genes is insufficient to allow annotation and use of the genomic information. A series of expressed sequence tags of 5' ends of five full-length enriched cDNA libraries (SUSFLECKs) were functionally characterized. SUSFLECKs were constructed from porcine abdominal fat, induced fat cells, loin muscle, liver, and pituitary gland, and were composed of non-normalized and normalized libraries. A total of 55,658 ESTs that were sequenced once from the 5′ ends of clones were produced and assembled into 17,684 unique sequences with 7,736 contigs and 9,948 singletons. In Gene Ontology analysis, two significant biological process leaf nodes were found: gluconeogenesis and translation elongation. In functional domain analysis based on the Pfam database, the beta transducin repeat domain of WD40 protein was the most frequently occurring domain. Twelve genes, including SLC25A6, EEF1G, EEF1A1, COX1, ACTA1, SLA, and ANXA2, were significantly more abundant in fat tissues than in loin muscle, liver, and pituitary gland in the SUSFLECKs. These characteristics of SUSFLECKs determined by EST analysis can provide important insight to discover the functional pathways in gene networks and to expand our understanding of energy metabolism in the pig.

A Study on the Profiles Transition and Storage Movement on the Profiles at HAEUNDAE Beach (해운대(海雲臺)사빈(砂濱)의 단면(斷面)의 천이주행(遷移走行) 및 저류(貯溜) 표사량(漂砂量) 변화특성(變化特性)에 관한 연구(研究))

  • Yang, Yun Mo;Ham, Gye Un
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.3 no.1
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    • pp.1-12
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    • 1983
  • The wave and hydrologic climate at a beach location are everchanging and coastal engineers have always been interested in the deformation of the natural beach caused by wave action over short or long interval of time. The drift of sand on a beach particulary manifests itself when blocked construction of seawalls, jetties, breakerwaters and groins etc.${\cdots}$ For this reason, the understanding and evaluation of littoral drift has been especially important to the coastal engineers. The resulting of the sand drift movement, such as the type of beach profile, width of beach, storage volume of the littoral sand over the profile are rapid everchange. We have studied the geological changes due to the littoral drift on a beach with field investigation and model tests in laboratory. But, it is impossible to make quantative correct analysis because of the factors are everchange and complicate. And then, most of study are incline to qualitative analysis. In this paper, authors studied mainly on the transition of beach profile and sediment storage on the profile using statistical field data as qualitative analytical method. The used theoretical beach transition model by Sonu and Beek have developed to obtain the change of HAEUNDAE beach backed with seawall. Results of this study indicate that the transition model are useful in the analysis of beach profile changement and the littoral drift movement on the beach. Qualitative analysises for HAEUNDAE beach are as follows. 1) Transition sequence of profile has 4 major transition for one cycle. 2) Storage sediment model of beach profile by Sonu and Beek well coincided with HAEUNDAE beach. 3) Seasonal cycle has ill-balanced process for the 5-yr. investigation.

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Construction of a Genetic Linkage Map in Radish(Raphanus sativus L.) Using RAPD Markers (RAPD 마커를 이용한 무의 유전자지도 작성)

  • Ahn, Choon-Hee;Choi, Su-Ryun;Lim, Yong-Pyo;Chung, Hae-Joon;Yae, Byeong-Woo;Yoon, Wha-Mo
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.151-159
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    • 2002
  • Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n=9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.