• Title/Summary/Keyword: detection technique

Search Result 4,102, Processing Time 0.046 seconds

The epigenetic phenotypes in transgenic Nicotiana benthamiana for CaMV 35S-GFP are mediated by spontaneous transgene silencing

  • Sohn, Seong-Han;Choi, Min-Sue;Kim, Kook-Hyung;Lomonossoff, George
    • Plant Biotechnology Reports
    • /
    • v.5 no.3
    • /
    • pp.273-281
    • /
    • 2011
  • Diverse epigenetic phenotypes are frequently found during research on transgenic plants. To understand the factors underlying such diversity, hundreds of independent 35S-GFP transgenic N. benthamiana plants were analyzed. The diverse GFP-expression phenotypes of the transgenic plants were classified into three major types based on the GFP expression patterns and their response to 35S-GFP agroinfiltration: steady-green, silenced and non-uniform phenotype. The non-uniform phenotype was further sub-divided into five minor phenotypes: variegated, red-dropped, on-silencing, partitioned and misty, according to the distribution of GFP expression on the leaves. Many of transgenic plants continuously generated diverse phenotypes over several generations despite the transgene identity. Such epigenetic GFP phenotyping was found to be the result of spontaneous transgene silencing mediated by either or both of post-transcriptional gene silencing (PTGS) and transcriptional gene silencing (TGS). This finding was verified by the detection of 21- and 24-nt small interfering RNA (siRNA) molecules, and DNA methylation in the transgenic plants that showed repeated epigenetic variation. Agroinfiltration demonstrated that irregular distribution of GFP on a leaf was the result of erratic transgene silencing, and the technique also proved to be a rapid and effective method for selecting fully silenced plants within 3 days. Furthermore, two novel phenotypes described are potential materials for in-depth investigations into the genes and mechanisms responsible for spontaneous transgene silencing.

Occurrence of Sesame Phyllody Disease in Korea and Detection of Its Phytoplasma (참깨 엽화병의 발생과 파이토플라스마의 검출)

  • Han, Mu-Seok;Noh, Eun-Woon;Yun, Jeong-Koo
    • Korean Journal Plant Pathology
    • /
    • v.13 no.4
    • /
    • pp.239-243
    • /
    • 1997
  • In August 1996, phyllody disease of sesame (Sesamum indicum L.) caused by phytoplasmas was observed at Boeun, Chungbuk Province, Korea. Symptoms included extreme proliferation of growing tips and numerous small leaves, giving the infected plant a witche's-broom effect. Parts or all of the floral parts were transformed into green leaf-like structures, and little or no seeds were produced. Transmission Electron microscopy revealed the presence of phytoplasmas in the phloem sieve elements of infected plant. Since the infected sesame plants were growing near by phytoplasma infected jujubes (Zizyphus jujubu), we tried a polymerase chain reaction (PCR) technique to identify these two causal phytoplasmas. The DNA extracted from the stems of infected sesame plant was PCR-amplified using a primer set specific to 16S rRNA gene of known phytoplasmas. The amplification generated a 1.4kb band in both sesame samples and phytoplasma-infected jujubes, which also suggests the sesame plants were infected with phytoplasmas. The restriction digestion of the amplified band by four different enzymes, AluI, HaeIII, HinfI or TaqI revealed that the phytoplasmas infecting jujubes and sesame plants were of different groups.

  • PDF

Fast and Accurate Determination of Algal Toxins in Water using Online Preconcentration and UPLC-Orbitrap Mass Spectrometry (온라인 시료주입과 UPLC-Orbitrap 질량분석법을 이용한 수질 조류독소의 고속분석방법 개발 및 환경시료적용)

  • Jang, Je-Heon;Kim, Yun-Seok;Choi, Jae-Won
    • Journal of Korean Society on Water Environment
    • /
    • v.28 no.6
    • /
    • pp.843-850
    • /
    • 2012
  • Due to the fast response to algae bloom issue in drinking water treatment plant, very fast determination methodology for algal toxin is required. In this study, column switching technique based online preconcentration method was combined with high resolution full scan mass spectrometer to save sample preparation time and to obtain fast and accurate result. After parameter optimization of online preconcentration, 1mL filtered sample was directly injected to trap column with switching valve system. Next, target toxins are eluted by 98% acetonitrile and analysed with 150 - 1,100 amu scan range at 50,000 resolving power. Method detection limit (MDL) for microcystin-LR, the most toxic isomer, was 0.1 ng/mL and others such as microcystin-YR, microcystin-RR and nodularin were 0.08, 0.03 and 0.04 ng/mL, respectively. This is the best improved sensitivities with 1mL volume in the literature. Furthermore, due to the use of ultra pressure HPLC (UPLC), the whole method run was completed in 4 min. Real sample applications for 173 sample including 55 surface water and 118 treatment plant samples for raw and treated water could be done within 16 hours. In our calculation, this methodology is roughly 80% faster than the previous manual solid-phase extraction with LC-MS/MS method.

Loop-mediated Isothermal Amplification (LAMP) for Detection of Streptococcus parauberis (Loop-mediated isothermal amplification (LAMP)법을 이용한 Streptococcus parauberis 의 신속 진단)

  • Moon, Kyung-Mi;Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
    • /
    • v.24 no.4
    • /
    • pp.428-436
    • /
    • 2014
  • Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

Study on Soot Primary Particle Size Measurement in Ethylene Diffusion Flame by Time-Resolved Laser-Induced Incandescence (시분해 레이저 유도 백열법을 이용한 에틸렌 확산 화염에서의 매연 일차입자크기 측정에 관한 연구)

  • Kim Gyu-Bo;Cho Seung-Wan;Lee Jong-Ho;Jeong Dong-Soo;Chang Young-June;Jeon Chung-Hwan
    • Transactions of the Korean Society of Mechanical Engineers B
    • /
    • v.30 no.10 s.253
    • /
    • pp.973-981
    • /
    • 2006
  • Recently there is an increasing interest in particulate matter emission because of new emission regulations, health awareness and environmental problems. It requires to improve particulate measurement techniques as well as to reduce soot emissions from combustion systems. As mentioned above, it is demanded that reduction techniques together with measurement techniques of exhausted particulate matters in combustion systems such as vehicles. However, measurement techniques of particulate matters should be prior to reduction techniques of that because it is able to know an increase and a decrease of exhausted particulate matters when measured particulate matters. Therefore, in this study, we report the measurement of soot primary-particle size using time-resolved laser induced incandescence (TIRE-LII) technique in laminar ethylene diffusion flame. As an optical method, laser induced incandescence is one of well known methods to get information for spatial and temporal soot volume fraction and soot primary particle size. Furthermore, TIRE-LII is able to measure soot primary particle size that is decided to solve the decay ate of signal S $(t_1)$ and S $(t_2)$ at two detection time. In laminar ethylene diffusion flame, visual flame height is 40 mm from burner tip and measurement points are height of 15, 20, 27.5, 30 mm above burner tip along radial direction. As increasing the height of the flame from burne. tip, primary particle size was increased to HAB(Height Above Burner tip)=20mm, and then decreased from HAB=27.5 mm to 30 mm. This results show the growth and oxidation processes for soot particles formed by combustion.

3D Building Reconstruction and Visualization by Clustering Airborne LiDAR Data and Roof Shape Analysis

  • Lee, Dong-Cheon;Jung, Hyung-Sup;Yom, Jae-Hong
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
    • /
    • v.25 no.6_1
    • /
    • pp.507-516
    • /
    • 2007
  • Segmentation and organization of the LiDAR (Light Detection and Ranging) data of the Earth's surface are difficult tasks because the captured LiDAR data are composed of irregularly distributed point clouds with lack of semantic information. The reason for this difficulty in processing LiDAR data is that the data provide huge amount of the spatial coordinates without topological and/or relational information among the points. This study introduces LiDAR data segmentation technique by utilizing histograms of the LiDAR height image data and analyzing roof shape for 3D reconstruction and visualization of the buildings. One of the advantages in utilizing LiDAR height image data is no registration required because the LiDAR data are geo-referenced and ortho-projected data. In consequence, measurements on the image provide absolute reference coordinates. The LiDAR image allows measurement of the initial building boundaries to estimate locations of the side walls and to form the planar surfaces which represent approximate building footprints. LiDAR points close to each side wall were grouped together then the least-square planar surface fitting with the segmented point clouds was performed to determine precise location of each wall of an building. Finally, roof shape analysis was performed by accumulated slopes along the profiles of the roof top. However, simulated LiDAR data were used for analyzing roof shape because buildings with various shapes of the roof do not exist in the test area. The proposed approach has been tested on the heavily built-up urban residential area. 3D digital vector map produced by digitizing complied aerial photographs was used to evaluate accuracy of the results. Experimental results show efficiency of the proposed methodology for 3D building reconstruction and large scale digital mapping especially for the urban area.

Fabrication and Sensing Capability of Cholesterol Sensors Based on ZnO Nanofibers (산화아연 나노섬유 기반 콜레스테롤 센서의 제작과 성능)

  • Jo, So Yeon;Kim, Ji Yeong;Kim, Sang Sub
    • Korean Journal of Materials Research
    • /
    • v.23 no.5
    • /
    • pp.281-285
    • /
    • 2013
  • In the present work, ZnO nanofibers were applied to electrode materials for the detection of cholesterol. ZnO nanofibers were synthesized using the electrospinning technique with zinc acetate as a precursor. Electrospinning-synthesized ZnO nanofibers were uniformly distributed by properly controlling the electrospinning parameters. After the calcination treatment, nanofibers of pure ZnO phase were synthesized. Then, these fibers were successfully placed on Au-coated glass substrates by dispersion of ZnO nanofibers in ethanol, dropping, and drying, in sequence. Cholesterol oxidase was then immobilized onto the surface of the ZnO nanofibers. To enhance the immobilization, Nafion was additionally applied. The sensing performances of the fabricated ZnO nanofibers-based sensors were analyzed by cyclic voltammetry in terms of cholesterol concentration ranging from 100 to 400 mg/dl. In the I-V curves, measured by cyclic voltammetry, the ZnO nanofiber-based sensor showed a proportional current behavior with cholesterol concentrations in phosphate buffered saline solution. The sensitivity was measured and found to be $30.7nA/mM{\cdot}cm^2$, which is comparable to the values reported in the literature. After not only optimizing the shape of the ZnO nanofibers but also improving the adhesion nature between the ZnO nanofibers and the Au conducting layer, these fibers can be a good candidate for electrode materials in devices used to detect low concentrations of cholesterol in blood.

Detection of porcine encephalomyocarditis virus by in situ hybridization (In situ hybridization에 의한 돼지 뇌심근염 바이러스의 검출)

  • Oh, Sang-hyeon;Park, Nam-yong;Chung, Ci-young;Cho, Kyoung-oh;Lee, Bong-joo;Park, Young-seok;Park, Hyung-seon
    • Korean Journal of Veterinary Research
    • /
    • v.39 no.1
    • /
    • pp.148-158
    • /
    • 1999
  • The purpose of this study was to establish a rapid, reliable diagnostic method detecting Encephalomyocarditis virus(EMCV) RNA in formalin-fixed, paraffin-embedded tissues of EMCV naturally infected pigs by cDNA probe of EMC $K_3$, the EMCV strain isolated from Korea. Using a biotin-labelled nick translated probe for the cDNA marker. We made up for some defects of radiolabeled method. In sits hybridization(ISH) technique, differently from the other nucleic acid hybridization methods, is able to detect the virus genome specifically in the state of the intact shapes of cells and/or tissues. We succeeded in performing the experiment to detect the EMCV within 1~2 hours using the $MicroProbe^{TM}$ capaillary action system. In this study, we observed highly specific positive signals of red color by staining the paraffin-embedded tissue sections of naturally EMCV-infected pig organs or tissues, including brain, heart, kidney and lacrimal gland with the Fast Red TR salt/Naphtol phosphate chromogen. The results suggested that this ISH method is considered as a highly sensitive and reliable tool for molecular biologic diagnosis of the EMC viral disease.

  • PDF

Exploratory research on ultra-long polymer optical fiber-based corrosion sensing for buried metal pipelines

  • Luo, Dong;Li, Yuanyuan;Yang, Hangzhou;Sun, Hao;Chen, Hongbin
    • Smart Structures and Systems
    • /
    • v.26 no.4
    • /
    • pp.507-520
    • /
    • 2020
  • In order to achieve effective corrosion monitoring of buried metal pipelines, a Novel nondestructive Testing (NDT) methodology using ultra-long (250 mm) Polymer Optical Fiber (POF) sensors coated with the Fe-C alloy film is proposed in this study. The theoretical principle is investigated to clarify the monitoring mechanism of this method, and the detailed fabrication process of this novel POF sensor is presented. To validate the feasibility of this novel POF sensor, exploratory research of the proposed method was performed using simulated corrosion tests. For simplicity, the geometric shape of the buried pipeline was simulated as a round hot-rolled plain steel bar. A thin nickel layer was applied as the inner plated layer, and the Fe-C alloy film was coated using an electroless plating technique to precisely control the thickness of the alloy film. In the end, systematic sensitivity analysis on corrosion severity was further performed with experimental studies on three sensors fabricated with different metal layer thicknesses of 25 ㎛, 30 ㎛ and 35 ㎛. The experimental observation demonstrated that the sensor coated with 25 ㎛ Fe-C alloy film presented the highest effectiveness with the corrosion sensitivity of 0.3364 mV/g at Δm = 9.32 × 10-4 g in Stage I and 0.0121 mV/g in Stage III. The research findings indicate that the detection accuracy of the novel POF sensor proposed in this study is satisfying. Moreover, the simple fabrication of the high-sensitivity sensor makes it cost-effective and suitable for the on-site corrosion monitoring of buried metal pipelines.

Quantitative Measurement of Soot concentration by Two-Wavelength Correction of Laser-Induced Incandescence Signals (2파장 보정 Laser-Induced Incandescence 법을 이용한 매연 농도 측정)

  • 정종수
    • Transactions of the Korean Society of Automotive Engineers
    • /
    • v.5 no.3
    • /
    • pp.54-65
    • /
    • 1997
  • To quantify the LII signals from soot particle of flames in diesel engine cylinder, a new method has been proposed for correcting LII signal attenuated by soot particles between the measuring point and the detector. It has been verified by an experiment on a laminar jet ethylene-air diffusion flame. Being proportional to the attenuation, the ratio of LII signal at two different detection wavelengths can be used to correct the measured LIIsignal and obtain the unattenuated LII signal, from which the soot volume fraction in the flame can be estimated. Both the 1064-nm and frequency-doubled 532-nm beams from the Nd : YAG laser are used. Single-shot, one-dimensional(1-D) line images are recorded on the intensified CCD camera, with the rectangular-profile laser beam using 1-mm-diameter pinhole. Two broadband optical interference filters having the center wavelengths of 647 nm and 400 nm respectively and a bandwidth of 10 nm are used. This two-wavelength correction has been applied to the ethylene-air coannular laminar diffusion flame, previously studied on soot formation by the laser extinction method in this laboratory. The results by the LII measurement technique and the conventional laser extinction method at the height of 40 nm above the jet exit agreed well with each other except around outside of the peaks of soot concentration, where the soot concentration was relatively high and resulting attenuation of the LII signal was large. The radial profile shape of soot concentration was not changed a lot, but the absolute value of the soot volume fraction around outside edge changed from 4ppm to 6.5 ppm at r=2.8mm after correction. This means that the attenuation of LII signal was approximately 40% at this point, which is higher than the average attenuation rate of this flame, 10~15%.

  • PDF