• Title/Summary/Keyword: transformation region

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Development of System-Wide Functional Analysis Platform for Pathogenicity Genes in Magnaporthe oryzae

  • Park, Sook-Young;Choi, Jaehyuk;Choi, Jaeyoung;Kim, Seongbeom;Jeon, Jongbum;Kwon, Seomun;Lee, Dayoung;Huh, Aram;Shin, Miho;Jung, Kyungyoung;Jeon, Junhyun;Kang, Chang Hyun;Kang, Seogchan;Lee, Yong-Hwan
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.9-9
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    • 2014
  • Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.

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Analysis of Co-registration Performance According to Geometric Processing Level of KOMPSAT-3/3A Reference Image (KOMPSAT-3/3A 기준영상의 기하품질에 따른 상호좌표등록 결과 분석)

  • Yun, Yerin;Kim, Taeheon;Oh, Jaehong;Han, Youkyung
    • Korean Journal of Remote Sensing
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    • v.37 no.2
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    • pp.221-232
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    • 2021
  • This study analyzed co-registration results according to the geometric processing level of reference image, which are Level 1R and Level 1G provided from KOMPSAT-3 and KOMPSAT-3A images. We performed co-registration using each Level 1R and Level 1G image as a reference image, and Level 1R image as a sensed image. For constructing the experimental dataset, seven Level 1R and 1G images of KOMPSAT-3 and KOMPSAT-3A acquired from Daejeon, South Korea, were used. To coarsely align the geometric position of the two images, SURF (Speeded-Up Robust Feature) and PC (Phase Correlation) methods were combined and then repeatedly applied to the overlapping region of the images. Then, we extracted tie-points using the SURF method from coarsely aligned images and performed fine co-registration through affine transformation and piecewise Linear transformation, respectively, constructed with the tie-points. As a result of the experiment, when Level 1G image was used as a reference image, a relatively large number of tie-points were extracted than Level 1R image. Also, in the case where the reference image is Level 1G image, the root mean square error of co-registration was 5 pixels less than the case of Level 1R image on average. We have shown from the experimental results that the co-registration performance can be affected by the geometric processing level related to the initial geometric relationship between the two images. Moreover, we confirmed that the better geometric quality of the reference image achieved the more stable co-registration performance.

A study for improvement of far-distance performance of a tunnel accident detection system by using an inverse perspective transformation (역 원근변환 기법을 이용한 터널 영상유고시스템의 원거리 감지 성능 향상에 관한 연구)

  • Lee, Kyu Beom;Shin, Hyu-Soung
    • Journal of Korean Tunnelling and Underground Space Association
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    • v.24 no.3
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    • pp.247-262
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    • 2022
  • In domestic tunnels, it is mandatory to install CCTVs in tunnels longer than 200 m which are also recommended by installation of a CCTV-based automatic accident detection system. In general, the CCTVs in the tunnel are installed at a low height as well as near by the moving vehicles due to the spatial limitation of tunnel structure, so a severe perspective effect takes place in the distance of installed CCTV and moving vehicles. Because of this effect, conventional CCTV-based accident detection systems in tunnel are known in general to be very hard to achieve the performance in detection of unexpected accidents such as stop or reversely moving vehicles, person on the road and fires, especially far from 100 m. Therefore, in this study, the region of interest is set up and a new concept of inverse perspective transformation technique is introduced. Since moving vehicles in the transformed image is enlarged proportionally to the distance from CCTV, it is possible to achieve consistency in object detection and identification of actual speed of moving vehicles in distance. To show this aspect, two datasets in the same conditions are composed with the original and the transformed images of CCTV in tunnel, respectively. A comparison of variation of appearance speed and size of moving vehicles in distance are made. Then, the performances of the object detection in distance are compared with respect to the both trained deep-learning models. As a result, the model case with the transformed images are able to achieve consistent performance in object and accident detections in distance even by 200 m.

Molecular Cloning of Bacillus stearothermophilus cdd Gene Encoding Thermostable Cytidine/Deoxycytidine Deaminase (Bacillus stearothermophilus 의 내열성 시티딘/디옥시시티딘 디아미나제를 코드하는 cdd 유전자의 클로닝)

  • Soo, Chang-Jong;Song, Bang-Ho;Kim, Jong-Guk;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.334-342
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    • 1989
  • The Bacillus stearothermophilus cdd gene encoding cytidine deaminase (cytidine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5) was isolated through shot gun cloning by oomplementation of an E. coli cdd mutation. Primarily 3.0 kbp of the exogenote was cloned into the Pstl site of pBR322 (pJSC101). By subsequent deletion and subcloning from the insert of pJSC101 with cdd$^+$ and tetracycline resistancy, about 1.35 kbp of the EcoRI$_1$/PstI$_2$ fragment containing the cdd gene was isolated as pJSC201. The minicell experiment revealed a molecular mass of 33,000 dalton for polypeptide from the cloned DNA fragment complementing the cdd gene. From the lacZ fusion of 550 bp fragment of the EcoRI$_1$/AuaI as a putative promoter region, the transcription direction of the cdd gene on pJSC201 is from EcoRI towards the PstI sites, When the cdd gene was expressed in B. subtilis ED4O (cdd$^-$, pyr$^-$) by transformation with the E. coli-B. subtilis shuttle vector, the gene expression occured more efficiently than in E. coli and the gene appears to be stably maintained in B. subtitis as well as in E. coli.

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Development of 2.5D Photon Dose Calculation Algorithm (2.5D 광자선 선량계산 알고리즘 개발)

  • 조병철;오도훈;배훈식
    • Progress in Medical Physics
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    • v.10 no.2
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    • pp.103-114
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    • 1999
  • In this study, as a preliminary study for developing a full 3D photon dose calculation algorithm, We developed 2.5D photon dose calculation algorithm by extending 2D calculation algorithm to allow non-coplanar configurations of photon beams. For this purpose, we defined the 3d patient coordinate system and the 3d beam coordinate system, which are appropriate to 3d treatment planning and dose calculation. and then, calculate a transformation matrix between them. For dose calculation, we extended 2d "Clarkson-Cunningham" model to 3d one, which can calculate wedge fields as well as regular and irregular fields on arbitrary plane. The simple Batho's power-law method was implemented as an inhomogeneity correction. We evaluated the accuracy of our dose model following procedures of AAPM TG#23; radiation treatment planning dosimetry verifications for 4MV of Varian Clinac-4. As results, PDDs (percent depth dose) of cubic fields, the accuracy of calculation are within 1% except buildup region, and $\pm$3% for irregular fields and wedge fields. And for 45$^{\circ}$ oblique incident beam, the deviations between measurements and calculations are within $\pm$4%. In the case of inhomogeneity correction, the calculation underestimate 7% at the lung/water boundary and overestimate 3% at the bone/water boundary. At the conclusions, we found out our model can predict dose with 5% accuracy at the general condition. we expect our model can be used as a tool for educational and research purpose.. purpose..

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Transformation of Rice Embryogenic Cells by Electroporation Mediated Plasmid Uptake into Protoplasts 1. Plant Regeneration from Electroporated Protoplasts of Rice (원형질체 내 Plasmid Electroporation에 의한 벼 배발생세포의 형질전환 1. 벼의 Electroporation 원형질체로부터 식물체 재분화)

  • 김명덕;최성진김준철
    • KSBB Journal
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    • v.10 no.1
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    • pp.23-29
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    • 1995
  • Calli were induced from leaf base region of germinated rice(Oryza sativa L. cv. Nakdong) with high frequency of up to 65% on LS medium supplemented with $2.5mg/{\ell}2$, 4-D in the dark at $27^{\circ}C$. Embryogenic calli of pale yellow, globular type were selected and used for the initiation of cell suspension cultures in AA2 liquid medium with $2mg/\ell$ 2,4-D, 0.2mg/$\ell$ kinetin arid $0.1mg/\ell$ GA3. Protoplasts were isolated from the embryogenic cell suspensions after 4 months of culture and then were electroporated with 400V/cm for 1 msec. Electroporated protoplasts divided with plating efficiency of 1.1% on PCM liquid medium supplemented with $2.5mg/\ell$ 2, 4-D, $0.1mg/\ell$ kinetin and 10mM proline. The protoplasts-derived microcalli were cultured on $0.2{\mu}m$ membrane fitter placed onto LS2.5 solid medium containing fine suspension cells as a feeder cells, for 2 weeks in the dark at $27^{\circ}C$. After an additional 2 weeks of culture under fluorescent light of $30{\pm}/3{\mu}E$.m^{-2}S^{-1}, yellow calli of 2mm diameter were transferred to regeneration medium. Shoots were produced from the green spot of protoplasts-derived calli and plants were regenerated form protoplast-derived green calli with frequencies of 11∼33%.

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Method of Measuring Color Difference Between Images using Corresponding Points and Histograms (대응점 및 히스토그램을 이용한 영상 간의 컬러 차이 측정 기법)

  • Hwang, Young-Bae;Kim, Je-Woo;Choi, Byeong-Ho
    • Journal of Broadcast Engineering
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    • v.17 no.2
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    • pp.305-315
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    • 2012
  • Color correction between two or multiple images is very crucial for the development of subsequent algorithms and stereoscopic 3D camera system. Even though various color correction methods are proposed recently, there are few methods for measuring the performance of these methods. In addition, when two images have view variation by camera positions, previous methods for the performance measurement may not be appropriate. In this paper, we propose a method of measuring color difference between corresponding images for color correction. This method finds matching points that have the same colors between two scenes to consider the view variation by correspondence searches. Then, we calculate statistics from neighbor regions of these matching points to measure color difference. From this approach, we can consider misalignment of corresponding points contrary to conventional geometric transformation by a single homography. To handle the case that matching points cannot cover the whole regions, we calculate statistics of color difference from the whole image regions. Finally, the color difference is computed by the weighted summation between correspondence based and the whole region based approaches. This weight is determined by calculating the ratio of occupying regions by correspondence based color comparison.

Field Observations of Spatial Structure of Hydrodynamics Including Waves and Currents in the Haeundae Coast (해운대의 파랑 및 흐름 구조의 특성파악을 위한 현장 관측실험)

  • Do, Kideok;Yoo, Jeseon;Lee, Hee Jun;Do, Jong-Dae;Jin, Jae-Youll
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.27 no.4
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    • pp.228-237
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    • 2015
  • Field observations were conducted to collect hydrodynamic and morphological data, which are needed to account for mechanisms of bathymetry changes caused by physical forcings, in Haeundae beach. In order to quantitatively describe characteristics of wave transformations and current patterns in space in winter and summer, in-situ sensors for measuring waves and current profiles were installed at three locations in the cross-shore direction and also three locations in the along-shore direction. As for the results of wave measurements, waves with main direction from the east dominate in winter while waves are incident from the S and the ESE in summer. Analysis of current data reveals that currents over the study domain are considerably influenced by a pattern of tidal motions, thereby, mainly oscillating in the direction of tidal currents, i.e., east-west directions, in both winter and summer. Currents tend to be influenced by local bathymetry in the shallow water region, with the direction changed along the depth contours and the magnitude reduced as they approach the shoreline. The results analysed from the hydrodynamic data through this study can be further combined with the morphological and bathymetry data, leading to the quantification of seasonal sediment transport rates and sand budget changes.

Effective Morphological Layer Segmentation Based on Edge Information for Screen Image Coding (스크린 이미지 부호화를 위한 에지 정보 기반의 효과적인 형태학적 레이어 분할)

  • Park, Sang-Hyo;Lee, Si-Woong
    • The Journal of the Korea Contents Association
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    • v.13 no.12
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    • pp.38-47
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    • 2013
  • An image coding based on MRC model, a kind of multi-layer image model, first segments a screen image into foreground, mask, and background layers, and then compresses each layer using a codec that is suitable to the layer. The mask layer defines the position of foreground regions such as textual and graphical contents. The colour signal of the foreground (background) region is saved in the foreground (background) layer. The mask layer which contains the segmentation result of foreground and background regions is of importance since its accuracy directly affects the overall coding performance of the codec. This paper proposes a new layer segmentation algorithm for the MRC based image coding. The proposed method extracts text pixels from the background using morphological top hat filtering. The application of white or black top hat transformation to local blocks is controlled by the information of relative brightness of text compared to the background. In the proposed method, the boundary information of text that is extracted from the edge map of the block is used for the robust decision on the relative brightness of text. Simulation results show that the proposed method is superior to the conventional methods.

In vitro endonucleolytic cleavage of synthesized cucumber mosaic virus RNA by hammerhead ribozyme (인공적으로 합성한 오이모자이크 바이러스 RNA의 헤머헤드 ribozyme에 의한 시험관내에서의 절단)

  • Park, Sang-Gyu;Hwang, Young-Soo
    • Applied Biological Chemistry
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    • v.37 no.1
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    • pp.56-63
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    • 1994
  • Oligonucleotides for a conserved region of the coat protein gene of cucumber mosaic virus (CMV) and a hammerhead structure ribozyme against CMV RNA were synthesized using a DNA synthesizer. Both strands of oligonucleotides were annealed and restricted with BamHI/SacI, then cloned into a plasmid pBS SK (+). The cloned CMV substrate and ribozyme were sequenced to verify correct constructions. In vitro transcriptions were carried out by using T7 RNA polymerase with BssHII or SspI digests of $1\;{\mu}g$ of substrate and ribozyme clones. The size of substrate RNA was 176 nucleotides (nt) containing 50 nt of CMV RNA sequence, 6 nt of XbaI restriction site and 120 nt of vector-derived sequence in the case of BssHII digest. The size of ribozyme RNA was 164 nt containing 40 nt of ribozyme RNA sequence and same sequences of substrate. Substrate RNA was efficiently cleaved into two fragments (96 nt and 80 nt) by ribozyme RNA. This endonucleolytic cleavage occurred more efficiently at $55^{\circ}C$ than $37^{\circ}C$. SspI digest-derived substrate RNA (2234 nt) was also cleaved into two fragments by the same ribozyme. SspI digest-derived ribozyme RNA (2222 nt) cleaved the above substrate to two fragments. In vitro-tested ribozyme construct is being cloned into a plant transformation vector to develop virus-resistant plants.

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