• Title/Summary/Keyword: GM-detection technology

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Event-specific Detection Methods for Genetically Modified Maize MIR604 Using Real-time PCR

  • Kim, Jae-Hwan;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1118-1123
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    • 2009
  • Event-specific real-time polymerase chain reaction (PCR) detection method for genetically modified (GM) maize MIR604 was developed based on integration junction sequences between the host plant genome and the integrated transgene. In this study, 2 primer pairs and probes were designed for specific amplification of 100 and 111 bp DNA fragments from the zSSIIb gene (the maize endogenous reference gene) and MIR604. The quantitative method was validated using 3 certified reference materials (CRMs) with levels of 0.1, 1, and 10% MIR604. The method was also assayed with 14 different plants and other GM maize. No amplification signal was observed in real-time PCR assays with any of the species tested other than MIR604 maize. As a result, the bias from the true value and the relative deviation for MIR604 was within the range from 0 to 9%. Precision, expressed as relative standard deviation (RSD), varied from 2.7 to 10% for MIR604. Limits of detections (LODs) of qualitative and quantitative methods were all 0.1%. These results indicated that the event-specific quantitative PCR detection system for MIR604 is accurate and useful.

Detection Method for Unapproved Genetically Modified Rose Plants in Korea Using Duplex Polymerase Chain Reaction (우리나라 미승인 유전자변형 장미의 duplex PCR검출법)

  • Kim, Jae-Hwan;Park, Young-Doo;Kim, Hae-Yeong
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.672-677
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    • 2010
  • A duplex PCR method was developed to detect a transformation vector pSPB130 used in the development of a genetically modified (GM) rose plant. To detect a GM rose plant, the anthocyanin synthase ($ANS$) was used as an endogenous reference gene of rose in PCR detection. The primer pair RHANS-KF/KR producing 107 bp amplicon was used to amplify the $ANS$ gene and no amplified product was observed in any of the 9 different plants used as a template. The primer pair GMRH-KF/KR was designed to amplify the junction sequence between 35S promoter and flavonoid 3',5'-hydroxylase ($F3^{\prime}5^{\prime}H$) gene in pSPB130. The detection limit of the duplex PCR method is approximately 0.5%. This result indicates that this duplex PCR method could be useful for monitoring unauthorized GM rose in Korea.

Production and Characterization of Monoclonal Antibodies Specific to PAT Protein Expressed in Genetically Modified Herbicide-Resistance Maize (제초제 내성 유전자 변형 옥수수 중 PAT단백질에 특이한 단크론성 항체의 생산과 특성 확인)

  • Kim, Sol-A;Lee, Jeong-Eun;Shim, Won-Bo;Kang, Sung-Jo;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.193-199
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    • 2018
  • In this study, PAT protein of genetically modified maize was prepared from the recombinant E. coli strain BL21 (DE3), and mice were immunized with the recombinant PAT protein. After cell fusion and cloning, two hybridoma cells (PATmAb-7 and PATmAb-12) were chosen since the monoclonal antibodies (Mabs) produced by them were confirmed to be specific to PAT protein in the indirect enzyme-linked immunsorbent assay (ELISA) and western blot tests. There were no cross-reactions of either Mabs to other GM proteins or to the extracts of non-GM maize. The ELISA based on the PATmAb-7 can sensitively detect 0.3 ng/g PAT protein in corn. These results indicate that the developed Mabs can be used as bio-receptors in the development of immunosensors and biosensors for the rapid and simple detection of GM corn adulterated in foods.

A Simple and Reliable Method for Preparation of Cross-Contamination-Free Plant Genomic DNA for PCR-Based Detection of Transgenes

  • Hwang, Seon-Kap;Kim, Young-Mi
    • BMB Reports
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    • v.33 no.6
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    • pp.537-540
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    • 2000
  • A simplified but reliable method was developed for the polymerase chain reaction (PCR)-based detection of genetically modified (GM) plants. The modified CTAB (mCTAB) method enabled us to prepare a high quality of genomic DNA from several hundred plant leaf samples in one day. Using DNA samples prepared from seven dicots and two monocots, approximately 1.75-kb regions spanning 17 S to 25 S ribosomal RNA genes were successfully amplified in a 2X PCR pre-mix containing BLOTTO. Further fidelity assessment of the mCTAB method by PCR analysis with Roundup Ready soybean (RRS) and non-RRS plants showed that the DNA samples prepared alternately from each of two lines were evidently free of cross-contamination. These results demonstrate that the mCTAB method is highly recommended for the rapid detection of transgenes in large numbers of leaf samples from diverse transgenic plants.

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Detection of Eight Different Events of Genetically Modified Maize by Multiplex PCR Method

  • Kim, Jae-Hwan;Song, Hee-Sung;Heo, Mun-Seok;Lee, Woo-Young;Lee, Soon-Ho;Park, Sun-Hee;Park, Hye-Kyung;Kim, Myung-Chul;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • v.15 no.1
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    • pp.148-151
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    • 2006
  • Multiplex PCR was performed to simultaneously detect eight different events of genetically modified (GM) maize. Specific primers were constructed from GA21, T25, TC1507, Mon810, Mon863, Event176, Bt11, and NK603 events of GM maize. Using this PCR method, specific GM maize was monitored in commercialized foods and feed.

A Tunable Band-Pass Filter for Multi Bio-Signal Detection (대역폭 조정 가능한 다중 생체 신호 처리용 대역 통과 필터 설계)

  • Jeong, Byeong-Ho;Lim, Shin-Il;Woo, Deok-Ha
    • Journal of IKEEE
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    • v.15 no.1
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    • pp.57-63
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    • 2011
  • This paper presents a tunable band pass filter (BPF) for multi bio-signal detection. The bandwidth can be controlled by the bias current of transconductance (gm), while conventional BPF exploited switchable capacitor array for band selection. With this design technique, the die area of proposed BPF reduced to at least one tenth the area of conventional design. The simulation results show the high cut-off frequency tuning range of from 100Hz to 1Khz. The circuit was implemented with a 0.18um CMOS standard technology. Total current consumption is 1uA at the supply voltage of 1V with sub-threshold design technique.

A detector system for searching lost γ-ray source

  • Khan, Waseem;He, Chaohui;Cao, Yu;Khan, Rashid;Yang, Weitao
    • Nuclear Engineering and Technology
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    • v.52 no.7
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    • pp.1524-1531
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    • 2020
  • The aim of this work is to develop a Geiger-Muller (GM) detector system for robot to look for a radioactive source in case of a nuclear emergency or in a high radiation environment. In order to find a radiation source easily, a detector system, including 3 detectors, was designed to search γ-ray radiation sources autonomously. First, based on GEANT4 simulation, radiation dose rates in 3 Geiger-Muller (GM) counters were simulated at different source-detector distances, distances between detectors and angles. Various sensitivity analyses were performed experimentally to verify the simulated designed detector system. A mono-energetic 137Cs γ-ray source with energy 662 keV and activity of 1.11 GBq was used for the observation. The simulated results were compared with the experimental dose rate values and good agreements were obtained for various cases. Only based on the dose rates in three detectors, the radiation source with a specific source activity and angle was localized in the different location. A method was adopted with the measured dose rates and differences of distances to find the actual location of the lost γ-ray source. The corresponding angles of deviation and detection limits were calculated to determine the sensitivity and abilities of our designed detector system. The proposed system can be used to locate radiation sources in low and high radiation environments.

Immunoblotting Assay for Glyphosate-tolerant Genetically Modified Soybean in Soybean Products (식품 중 제초제내성 유전자재조합 콩의 immunoblotting 분석)

  • Son, Dae-Yeul;Moon, Jung-Hee;Ahn, Kang-Mo;Shon, Dong-Hwa;Lee, Kwang-Shin;Sim, Hee-Young;Han, Young-Shin;Lee, Sang-Il
    • Korean Journal of Food Science and Technology
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    • v.36 no.3
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    • pp.369-374
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    • 2004
  • Genetically modified (GM) soybean Roundup Ready carries Agrobacterium sp. CP4 gene, which expresses 5-enolpyruvylshikimate-3-phosphate synthase (CP4EPSPS). CP4EPSPS in GM soybeans and soybean curds was screened using CP4EPSPS-specific polyclonal and monoclonal antibodies (pab and mab, respectively) by immunoblotting. Isolated recombinant CP4EPSPS was detected at detection limits of $0.006\;and\;0.0006{\mu}g$, whereas those of CP4EPSPS expressed in GM soybean were $0.001\;and\;0.0001{\mu}g$g, using mab and pab, respectively. From nine screened soybean curds, two had positive results with pab Immunoblotting method with pab and mab developed in this study could be applied to screen glyphosate-tolerant GM soybeans in soybean products.

A New Concept for Efficient Sensitivity Amplification of a QCM Based Immunosensor for TNF-α by Using Modified Magnetic Particles under Applied Magnetic Field

  • Bahk, Yeon-Kyoung;Kim, Hyung-Hoon;Park, Deog-Su;Chang, Seung-Cheol;Go, Jeung-Sang
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4215-4220
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    • 2011
  • This study introduces a new concept for a simple, efficient and cheap sensitivity amplification of a Quartz Crystal Microbalance (QCM) based immunosensor system for the detection of tumor necrosis factor-alpha (TNF-${\alpha}$, TNF) by using an in-built magnetic system. The frequency shift due to the applied magnetic field was successfully observed on magnetic particles labeled detection antibodies, anti-human TNF-${\alpha}$, which were bound to the immunologically captured TNF-${\alpha}$ on the gold coated quartz crystals. In the present system, the magnitude of frequency shift depends on both the strength of magnetic field and the amount of target antigen applied. Significant signal amplification was observed when the additional built-in residual stress generated by the modified magnetic particles under the magnetic field applied. Used in conjunction with a sandwich type non-competitive immunoassay format, the lower detection limit was calculated to be 25 $ngmL^{-1}$ and showed good linearity up to TNF-${\alpha}$ concentrations as high as 2.0 ${\mu}gmL^{-1}$. The sensitivity, most importantly, was improved up to 4.3 times compared with the same QCM system which was used only an antigen-antibody binding without additional magnetic amplification.

Quantitation of CP4 5-Enolpyruvylshikimate-3-Phosphate Synthase in Soybean by Two-Dimensional Gel Electrophoresis

  • KIM YEON-HEE;CHOI SEUNG JUN;LEE HYUN-AH;MOON TAE WHA
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.25-31
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    • 2006
  • Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.