• 제목/요약/키워드: STE

검색결과 105건 처리시간 0.025초

Genetic Transformation of Geobacillus kaustophilus HTA426 by Conjugative Transfer of Host-Mimicking Plasmids

  • Suzuki, Hirokazu;Yoshida, Ken-Ichi
    • Journal of Microbiology and Biotechnology
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    • 제22권9호
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    • pp.1279-1287
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    • 2012
  • We established an efficient transformation method for thermophile Geobacillus kaustophilus HTA426 using conjugative transfer from Escherichia coli of host-mimicking plasmids that imitate DNA methylation of strain HTA426 to circumvent its DNA restriction barriers. Two conjugative plasmids, pSTE33T and pUCG18T, capable of shuttling between E. coli and Geobacillus spp., were constructed. The plasmids were first introduced into E. coli BR408, which expressed one inherent DNA methylase gene (dam) and two heterologous methylase genes from strain HTA426 (GK1380-GK1381 and GK0343-GK0344). The plasmids were then directly transferred from E. coli cells to strain HTA426 by conjugative transfer using pUB307 or pRK2013 as a helper plasmid. pUCG18T was introduced very efficiently (transfer efficiency, $10^{-5}-10^{-3}\;recipient^{-1}$). pSTE33T showed lower efficiency ($10^{-7}-10^{-6}\;recipient^{-1}$) but had a high copy number and high segregational stability. Methylase genes in the donor substantially affected the transfer efficiency, demonstrating that the host-mimicking strategy contributes to efficient transformation. The transformation method, along with the two distinguishing plasmids, increases the potential of G. kaustophilus HTA426 as a thermophilic host to be used in various applications and as a model for biological studies of this genus. Our results also demonstrate that conjugative transfer is a promising approach for introducing exogenous DNA into thermophiles.

Surface Ozone Episode Due to Stratosphere-Troposphere Exchange and Free Troposphere-Boundary Layer Exchange in Busan During Asian Dust Events

  • Moon, Y.S.;Kim, Y.K.;K. Strong;Kim, S.H.;Lim, Y.K.;Oh, I.B.;Song, S.K.
    • 한국환경과학회지
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    • 제11권5호
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    • pp.419-436
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    • 2002
  • The current paper reports on the enhancement of O$_3$, CO, NO$_2$, and aerosols during the Asian dust event that occurred over Korea on 1 May 1999. To confirm the origin and net flux of the O$_3$, CO, NO$_2$, and aerosols, the meteorological parameters of the weather conditions were investigated using Mesoscale Meteorological Model 5(MM5) and the TOMS total ozone and aerosol index, the back trajectory was identified using the Hybrid Single-Particle Lagrangian Integrated Trajectory Model(HYSPLIT), and the ozone and ozone precursor concentrations were determined using the Urban Ashed Model(UAM). In the presence of sufficiently large concentrations of NO$\sub$x/, the oxidation of CO led to O$_3$ formation with OH, HO$_2$, NO, and NO$_2$ acting as catalysts. The sudden enhancement of O$_3$, CO, NO$_2$ and aerosols was also found to be associated with a deepening cut-off low connected with a surface cyclone and surface anticyclone located to the south of Korea during the Asian dust event. The wave pattern of the upper trough/cut-off low and total ozone level remained stationary when they came into contact with a surface cyclone during the Asian dust event. A typical example of a stratosphere-troposphere exchange(STE) of ozone was demonstrated by tropopause folding due to the jet stream. As such, the secondary maxima of ozone above 80 ppbv that occurred at night in Busan, Korea on 1 May 2001 were considered to result from vertical mixing and advection from a free troposphere-boundary layer exchange in connection with an STE in the upper troposphere. Whereas the sudden enhancement of ozone above 100 ppbv during the day was explained by the catalytic reaction of ozone precursors and transport of ozone from a slow-moving anticyclone area that included a high level of ozone and its precursors coming from China to the south of Korea. The aerosols identified in the free troposphere over Busan, Korea on 1 May 1999 originated from the Taklamakan and Gobi deserts across the Yellow River. In particular, the 1000m profile indicated that the source of the air parcels was from an anticyclone located to the south of Korea. The net flux due to the first invasion of ozone between 0000 LST and 0600 LST on 1 May 1999 agreed with the observed ground-based background concentration of ozone. From 0600 LST to 1200 LST, the net flux of the second invasion of ozone was twice as much as the day before. In this case, a change in the horizontal wind direction may have been responsible for the ozone increase.

Intestinal Helminths Recovered from Humans in Xieng Khouang Province, Lao PDR with a Particular Note on Haplorchis pumilio Infection

  • Chai, Jong-Yil;Sohn, Woon-Mok;Jung, Bong-Kwang;Yong, Tai-Soon;Eom, Keeseon S.;Min, Duk-Young;Insisiengmay, Bounnaloth;Insisiengmay, Sithat;Phommasack, Bounlay;Rim, Han-Jong
    • Parasites, Hosts and Diseases
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    • 제53권4호
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    • pp.439-445
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    • 2015
  • A survey of intestinal helminths was undertaken in riparian people in Xieng Khouang Province, Lao PDR. Fecal specimens were collected from 643 people (289 males and 354 females) residing in 4 districts (Nonghet, Kham, Phoukout, and Pek) and were examined by the Kato-Katz technique. The overall helminth egg positive rate was 41.2%, and hookworms revealed the highest prevalence (32.7%) followed by Trichuris trichiura (7.3%) and Ascaris lumbricoides (5.6%). The positive rate for small trematode eggs (STE), which may include Opisthorchis viverrini, heterophyids, and lecithodendriids, was 4.4%. For recovery of adult helminths, 12 STE or nematode/cestode egg-positive people were treated with 40 mg/kg praziquantel and 15 mg/kg pyrantel pamoate, and then purged. Mixed infections with 2 Haplorchis species (H. pumilio and H. taichui), Centrocestus formosanus, Opisthorchis viverrini, a species of cestode (Taenia saginata), and several species of nematodes including hookworms and Enterobius vermicularis were detected. The worm load for trematodes was the highest for H. pumilio with an average of 283.5 specimens per infected person followed by C. formosanus, H. taichui, and O. viverrini. The worm load for nematodes was the highest for hookworms (21.5/infected case) followed by E. vermicularis (3.2/infected case). The results revealed that the surveyed areas of Xieng Khouang Province, Lao PDR are endemic areas of various species of intestinal helminths. The STE found in the surveyed population were verified to be those of heterophyids, particularly H. pumilio.

Subclinical left ventricular dysfunction in children after hematopoietic stem cell transplantation for severe aplastic anemia: a case control study using speckle tracking echocardiography

  • Kim, Beom Joon;Moon, Kyung Pil;Yoon, Ji-Hong;Lee, Eun-Jung;Lee, Jae Young;Kim, Seong Koo;Lee, Jae Wook;Chung, Nack Gyun;Cho, Bin;Kim, Hack Ki
    • Clinical and Experimental Pediatrics
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    • 제59권4호
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    • pp.190-195
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    • 2016
  • Purpose: Severe aplastic anemia (SAA), a fatal disease, requires multiple transfusion, immunosuppressive therapy, and finally, hematopoietic stem cell transplantation (HSCT) as the definitive treatment. We hypothesized that iron overloading associated with multiple transfusions and HSCT-related complications may adversely affect cardiac function. Left ventricular (LV) function was assessed in children after HSCT for SAA. Methods: Forty-six consecutive patients with a median age of 9.8 years (range, 1.5-18 years), who received HSCT for SAA and who underwent comprehensive echocardiography before and after HSCT, were included in this study. The data of LV functional parameters obtained using conventional echocardiography, tissue Doppler imaging (TDI), and speckle-tracking echocardiography (STE) were collected from pre- and post-HSCT echocardiography. These data were compared to those of 40 age-matched normal controls. Results: In patients, the LV ejection fraction, shortening fraction, end-diastolic dimension, mitral early diastolic E velocity, TDI mitral septal E' velocity, and STE LV longitudinal systolic strain rate (SSR) decreased significantly after HSCT. Compared to normal controls, patients had significantly lower post-HSCT early diastolic E velocity and E/A ratio. On STE, patients had significantly decreased LV deformational parameters including LV longitudinal systolic strain (SS), SSR, and diastolic SR (DSR), and circumferential SS and DSR. Serum ferritin levels showed weak but significant correlations (P<0.05) with LV longitudinal SS and SSR and circumferential SS and DSR. Conclusion: Subclinical LV dysfunction is evident in patients after HSCT for SAA, and was associated with increased iron load. Serial monitoring of cardiac function is mandatory in this population.

유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색 (Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato)

  • 신원선;김명희
    • 한국식품과학회지
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    • 제35권4호
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    • pp.591-597
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    • 2003
  • 국내에서 시판되는 감자와 수입 감자스낵류로부터 상용 DNA 추출 kit 및 CTAB-phenol/chloroform 추출법등을 이용하여 시료특성에 따른 genomic DNA를 추출방법을 선정하고 PCR 정성검사를 실시하였다. 생감자의 경우 STE 용액으로 과량의 전분을 제거한 다음 DNA를 추출한 경우 순도 높은 DNA를 추출할 수 있었으며 상용 추출 kit를 이용한 경우 lysis buffer와 함께 ${\alpha}-/{\beta}$-amylase를 각각 또는 혼합으로 처리하거나 추출된 DNA 용액에 마지막 단계에서 효소를 처리한 시료군에서 고순도의 DNA를 추출할 수 있었으며, 효소 처리군에서는 ${\alpha}-/{\beta}$-amylase를 혼합으로 처리한 경우에 DNA 추출수율이 높았다. 냉동가공감자의 경우 silica-coated bead법을 이용하여 효소를 처리한 경우와 CTAB-페놀 클로로포름 처리군에서 DNA가 추출되었다. 또한, 각 방법으로 추출한 DNA에 대하여 감자의 내인성 유전자인 Pss 프라이머를 사용하여 PCR을 한 결과 모든 시료에서 추출된 DNA에 대하여 내부표준유전자 증폭산물이 검출되었다. 고도의 가공처리를 거친 수입 감자스낵(fabricated potato chips)과 냉동가공 감자(frozen fried potato) 등은 계면활성제인 CTAB(cetyl trimethyl ammonium bromide)과 페놀-클로로포름 혼합액을 이용하여 추출하고 이를 template로 하여 PCR 증폭을 실시하였다. 그 결과, Fig. 8에 제시한 바대로 감자의 내인성 유전자인 Pss 특이적 산물인 216bp의 산물이 냉동감자가공품과 감자칩에서 검출되었으며 재조합유전자인 New leaf plus 유래의 증폭산물(234bp)와 New lear Y유래의 증폭산물(225bp)는 검출되지 않았다. 본 실험의 결과 시료의 가공특성과 적용한 추출 kit 및 방법에 따라 genomic DNA 순도 및 추출수율이 크게 차이가 났으며 이것이 결국 PCR 결과에 의음성 혹은 의양성 등에 영향을 미치게 될 것으로 판단된다. 또한, 동일한 DNA추출방법에 의해서도 DNA가 추출되지 않은 경우가 있어서 동일한 시료에서 2회 반복 추출하는 것이 의음성결과를 피할 수 있는 방법으로 판단된다.

Bone set-up을 통한 character animation Nothing제작을 통한 3D Animation 몇 가지 제안(提案)

  • 황길남
    • 만화애니메이션 연구
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    • 통권5호
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    • pp.531-534
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    • 2001
  • 창의적인 표현 이전에 무엇을 인식(認識)해야 하는가! 쉽게 생각하는 캐릭터모델링과 셋업(STE-UP) 과정이 몇 가지 옵션(Option)에 의하여 생성되는 것은 아니다. 애니메이션 제작에 따른 캐릭터 설정 배경, 인체구조와 모델링과의 관계, 인체구조와 애니메이션의 연관성 등에 대한 인식의 이해를 통해야 한다. 이러한 인식 속에서 3D 캐릭터애니메이션셋업(SET-UP)을 위한 기초인식과 기술을 통하여 원칙적인 이해에 근거하여 효율적, 효과적인 3D애니메이션 제작을 하기 위한 몇 가지 제안(提案)을 한다.

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NC 공정기계에서 온더머신 오차측정 및 보상 (On the machine error measurement and compensation)

  • 신동수;정성종
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 1992년도 한국자동제어학술회의논문집(국내학술편); KOEX, Seoul; 19-21 Oct. 1992
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    • pp.1096-1101
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    • 1992
  • In order tominimize fixing error of workpieces, circle, prismatic, sphere, cylindrical and sculptures types. Modification Rule by Indexing Table and Modification Rule by NC Program are developed for machining centers by using touch trigger probes. The Modification Rule by Indexing Table meas the alignment of workpiece to NC program through degree of freedoms of indexing table. The Modification Rule by NC Program is the alognment of NC program to workpiece ste-tp condition via the generation of NC progarm. A postprocessing module is alos developed for generating NC-part program(User Macro) to compensate for machining errors in end milling and boring processes. Developed method are verified by experiments.

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Development of a Highly Active Fluorescence-Based Detector for Yeast G Protein-Coupled Receptor Ste2p

  • Hong, Jin Woo;Ahn, Hee Jun;Baek, Jee Su;Hong, Eun young;Jin, Dong Hoon;Khang, Yong Ho;Hong, Nam Joo
    • Journal of Microbiology and Biotechnology
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    • 제28권10호
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    • pp.1589-1603
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    • 2018
  • Twenty analogs of $[Orn^6,D-Ala^9]{\alpha}-factor$ were synthesized and assayed for their biological activities: seven analogs of $[Orn^6,X^9]{\alpha}-factor$, seven analogs of $[X^6,D-Ala^9]{\alpha}-factor$, five analogs of $[X^5,X^6,D-Ala^9]{\alpha}-factor$, and native ${\alpha}-factor$ (X = amino acids). Their biological activities (halo, gene induction, and affinity) were measured using S. cerevisiae Y7925 and LM102 and compared with those of native ${\alpha}-factor$ (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. $[Dap^6,D-Ala^9]{\alpha}-factor$ with weak halo activity (10%) showed the highest receptor affinity (> 230%) and the highest gene induction activity (167%). $[Arg^6,D-Ala^9]{\alpha}-factor$ showed the highest halo activity (2,000%). The number of active binding sites per cell (about 20,000 for strain LM102) was determined using a newly-designed fluorescence-based detector, $[Arg^6,D-Ala^9]{\alpha}-factor-Edan$, with high sensitivity (12,500-fold higher than the absorption-based detector $[Orn^6]{\alpha}-factor-[Cys]_3$).

Improvement of RT-PCR Sensitivity for Fruit Tree Viruses by Small-scale dsRNA Extraction and Sodium Sulfite

  • Lee, Sin-Ho;Kim, Hyun-Ran;Kim, Jae-Hyun;Kim, Jeong-Soo
    • The Plant Pathology Journal
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    • 제20권2호
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    • pp.142-146
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    • 2004
  • Woody plant tissues contain great amounts of phenolic compounds and polysaccharides. These substances inhibit the activation of reverse transcriptase and/or Taq polymerase in RT-PCR. The commonly used multiple-step protocols using several additives to diminish polyphenolic compounds during nucleic acid extraction are time consuming and laborious. In this study, sodium sulfite was evaluated as an additive for nucleic acid extraction from woody plants and the efficiency of RT-PCR assay of commercial nucleic acid extraction kits and small-scale dsRNA extraction was compared. Sodium sulfite was used as an inhibitor against polyphenolic oxidases and its effects were compared in RNA extraction by commercial extraction kit and small-scale double-stranded RNA (dsRNA) extraction method for RT-PCR. During nucleic acid extraction, addition of 0.5%-1.5%(w/v) of sodium sulfite to lysis buffer or STE buffer resulted in lighter browning by oxidation than extracts without sodium sulfite and improved the RT-PCR detection. When commercial RNA extraction kit was used, optimal concentrations of sodium sulfite were variable according to the tested plant. However, with dsRNA as RT-PCR template, sodium sulfite 1.5% in STE buffer improved the detection efficiency of Apple chlorotic leaf spot virus (ACLSV) and Apple stem grooving virus (ASGV) in fruit trees, and reduced the unspecific amplifications signi-ficantly. Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

경주 감은사지서삼층석탑 충전제 선정과 보존처리 (Selection and Conservation for the Filler of Three-storied Stone Pagoda at the West of Gameunsaji Site in Gyeongju)

  • 이태종;김사덕;갈서연
    • 보존과학회지
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    • 제26권4호
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    • pp.361-370
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    • 2010
  • Gameunsaji West Stone Pagoda;Filler;Consolidation;Pore;Injection;Materials; 석조문화재의 내부공극은 강화처리만으로 효과를 보기 어렵고 고분자합성수지를 이용한 충전의 경우 재료적 이질감과 높은 강도, 저점도에 따른 유동성 향상으로 내부벽면만 접착, 강화되는 단점이 있다. 따라서 본 연구는 무기질 바탕의 다양한 충전제를 검색하여 충전효과검증을 통해 감은사지삼층석탑 공극부 충전에 효과적인 충전제를 선정, 보존처리를 실시하였다. 충전실험 결과 부재와 동일하거나 매우 유사한 물성을 지닌 KSE 500 STE + KSE Filler A KSE Filler B 조합을 사용하여 충전하였다. 서탑 전체표면적은 $252.6m^2$이며 이중 내부공극이 발달한 부분은 $17.77m^2$(7.03%)로 내부를 충전($24,885m{\ell}$)하였다.