• Title/Summary/Keyword: GM plant

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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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Identification of a host range determinant from Ralstonia solancearum race 3

  • Yeonhwa Jeong;Lee, Seungdon;Ingyu Hwang
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.71.2-71
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    • 2003
  • Ralstonia solancearum infects many solanaceous plants, however race 3 infects only potato and tomato weakly. To identify genes responsible for race specificity of R. solanacearum, we mobilized genomic library of LSD2029 (race 3) into LSD341 (race 1) and inoculated 1,000 transconjugants into hot pepper. One transconjugant that did not induce wilt symptom in hot pepper was isolated. We found that a cosmid clone, pRSl, conferred avirulence to LSD341. By deletion and mutational analyses of pRSl, we found the 0.9-kb PstI/Hindlll fragment carries avirulence functions. We sequenced the fragment and identified one possible open reading frame, a rsal gene, possibly encoding 110 amino acids. The rsal was preceded with a plant-inducible promoter (PIP) box, indicating that the gene might be regulated by HrpB. Interestingly, the promoter region of the rsal homolog in the strain GM11000 (race 1) did not have the PIP box. Rsal did not show any significant homologies with proteins in the database, indicating th e protein is different from the previously reported avirulence proteins. When we mutated the rsal gene by marker-exchange in LSD2029, the mutant was less virulent in potato.

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Identification of Introduced Gene and Its Expression and Gene Stability Assessment for Event Selection of Genetically Modified Plant toward Approval: Cucumber Mosaic Virus Resistant Hot Pepper (상업용 유전자 변형작물 이벤트 선발을 위한 도입유전자 확인, 발현 및 세대간 안정성 평가 : 오이모자이크바이러스 저항성 GM 고추)

  • Kang, Seung-Won;Han, Bal-Kum;Lee, Tae-Ho;Kim, Eun-Ji;Lee, Gung-Pyo
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.192-200
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    • 2012
  • For the development of genetically modified plants, it is important to verify various factors which potentially affect the risk assessment as well as to establish an experimental program to produce scientific and reliable data. However, it is a time and cost consuming process to develop GM plants as well as to prepare scientific and convincible data for government's approval. Therefore, using the transgenic hot pepper tolerant to a new CMV pathotype, we attempted to suggest few methodological procedures, such as probe saturation for southern blot analysis and RT-PCR and ELISA for expression analysis, for identification and stability evaluation of inserted gene in genetically modified plant which are required for submission for approval. Ten partially overlapped probes covering full length of inserted gene were produced. We could identify that the inserted gene was stacked as a single copy as well as no partial element existed. Also, we could identify the stability of the inserted gene stacked in hot pepper using probe saturation. In the expression analysis with RT-PCR and ELISA, we also could provide the stable expression of transcript and proteins in leaves and placenta and pericarp of fruits of the CMV-resistant hot pepper.

The Bacteriological Quality of Seawater in Kamak Bay, Korea (가막만해역 해수의 세균학적 수질 평가)

  • Kwon, Ji Young;Park, Kunbawui;Song, Ki Cheol;Oh, Eun Gyoung;Lee, Hee Jung;Jo, Mi-Ra;Kim, Ji Hoe;Son, Kwang Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.5
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    • pp.460-464
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    • 2012
  • From 1998 to 2008, the bacteriological quality of seawater in Kamak Bay, Korea, was monitored. The total and fecal coliforms in seawater were monitored as bacterial indicators, and the level of fecal coliform contamination was used to evaluate seawater quality. In the southern shellfish growing area of Kamak Bay, the observed fecal coliform geometric mean (GM) and estimated 90th percentile did not exceed the quality standards for fecal coliforms in seawater in Korea and th USA (GM<14 MPN/100 mL, 90th percentile<43 MPN/100 mL). The GM and estimated 90th percentile of the fecal coliform levels in the adjacent northern area of Kamak Bay were higher than in the southern area. Stations near wastewater discharge sites and urban areas in the north had the lowest quality and did not meet the bacteriological seawater quality criteria. The high fecal coliform levels in the Seonso coastal region near wastewater discharges and northern urban areas declined significantly following dredging and operation of a sewage treatment plant.

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.

In vitro and In vivo Effects of Gelidium amansii on Intestinal Immune System

  • Jun, Woo-Jin;Kim, Se-Han;Lee, Dae-Hee;Chun, Jin-Woong;Sim, Sang-In;Lee, Kwang-Won;Cho, Hong-Yon;Hong, Bum-Shik
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.147-151
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    • 2005
  • Purified compound with intestinal immune system-modulating properties, GWE-2c, was isolated from methanol extract of Gelidium amansii by sequential procedures with silica gel column, LH-20 Sephadex gel column, and thin-layer chromatographies. In the presence of GWE-2c, strong immunoactivity in Peyers patch cell-mediated bone marrow cells was observed in vitro. In vivo intestinal immune-modulating activity was also enhanced by crude phenolic compound (GWE) of G. amansii in a dose-dependent manner. Investigation of production of several cytokines in Peyer's patch cells upon stimulation with GWE in vivo revealed the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-6 increased. Results suggest that the phenolic compound from G. amansii represents immunopotentiator and biological response modifier at in vitro and in vivo levels.

Cre-Lox: A Tool for Removal of Marker Genes to Make GM Foods Safe

  • Zargar, Sajad M.;Mushtaq, Roohi;Joshi, Manisha;Prasad, D. Theertha;Bhat, Nazir Ahmad;Agrawal, Ganesh Kumar;Rakwal, Randeep
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.73-78
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    • 2007
  • The green revolution has significantly helped in increasing the food production. So far, various breeding methods have been exploited, besides them recombination DNA technology provides another approach for increasing the food production. By means of this technology the losses in food production incurred by various biotic and abiotic stresses can be effectively controlled. In most of the transgenic studies scientists have used antibiotic resistant genes as markers for easy selection of transformants but there are risks involved in use of GM foods. To make such foods safer and environment friendly we have discussed a novel strategy i.e. Cre-lox which involves site specific recombination. By means of Cre-lox the marker genes can be specifically removed once the selection of transformants is over. In addition, this strategy can be used to module the hybrid chromosomes, avoid gene silencing and incorporate single copy of a transgene for its higher expression.

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Development and Evaluation of a Mobile Environmental Radiation Measurement System That Can Switch between Low and High Dose Measurement Sections (저선량과 고선량 측정구간 변환이 가능한 모바일 방사선 측정시스템 개발 및 평가)

  • Lee, Hong-Yeon;Han, Sang-Jun;Kim, Bo-Gil;Lee, Geon-Ju;Kim, Seok-Hyeon;Kim, Jeong-Hun
    • Journal of radiological science and technology
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    • v.45 no.1
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    • pp.49-55
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    • 2022
  • This study is to develop a mobile type environmental radiation measurement system for emergency response or environmental radiation monitoring of local governments near nuclear facilities. A mobile radiation measurement system can monitor radiation by field beyond the spatial constraints of a fixed environmental radiation monitor. If installed in local government infrastructure such as public transportation, environmental radiation can be monitored without additional manpower and measurement work. In addition, it is designed to enable monitoring and measurement of radiation from low to high doses as well as the environment in preparation for radioactive disasters such as nuclear power plant accidents. It is expected that this system will be utilized not only in normal times but also in the event of a radiation accident to improve the disaster prevention capabilities of local governments.

Comparison of the Efficiency from Raw and Processed Corns by Five Different DNA Extraction Methods (다섯 가지 DNA 추출방법에 의한 옥수수 원료 및 가공시료의 DNA 추출 효율의 비교)

  • Lee, Hun-Hee;Song, Hee-Sung;Kim, Jae-Hwan;Lee, Woo-Young;Lee, Soon-Ho;Park, Sun-Hee;Park, Hye-Kyung;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.331-334
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    • 2005
  • In this study, the effects of five extraction methods for raw and processed corns were compared with respect to the integrity, yields and quality of DNA extracted from them and the results were assessed by PCR analysis. From the comparison of five extraction methods, DNA integrity showed a similar pattern. Amounts of genomic DNA obtained from the five extraction methods varies from $0.25{\mu}g\;to\;234{\mu}g$ per 1 g sample. The DNA yield extracted with CTAB method and DNeasy Plant Maxi kit is greater than that obtained from other extraction methods. These results would be applicable for the selection of an adequate extraction method for specific samples.

Risk assessment of genetically engineered rice Bt-9 resistant to Cnaphalocrocis medinalis: influence on above-ground arthropods in Korea

  • Oh, Sung-Dug;Bae, Eun Ji;Park, Soo-Yun;Lee, Bumkyu;Yun, Do Won;Suh, Sang Jae
    • Korean Journal of Agricultural Science
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    • v.46 no.4
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    • pp.827-841
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    • 2019
  • The effect of genetically engineered rice Bt-9 on the diversity and abundance of plant-dwelling insects and spiders was tested under field conditions. Genetically engineered rice Bt-9, expressing mCry1Ac1 from Bacillus thuringiensis, confers resistance to rice leaf roller (Cnaphalocrocis medinalis) and provides tolerance to the herbicide glufosinate (PPT). The study compared Bt-9 and two non-GM reference varieties, Ilmi-byeo and Dongjin-byeo, at LMO isolated fields in Gunwi (Kyungpook National University) and Jeonju (National Institute Agricultural Sciences) in Southern Korea in 2016 - 2017. A total of 40,817 individuals from 62 families and 11 orders were collected from the two living modified organism (LMO) isolated fields. From the three types of rice fields, a total of 13,982, 14,105, and 12,730 individuals from the Bt-9, Ilmi-byeo and Dongjin-byeo were collected, respectively. Throughout the study, the analysis of variance indicated no significant differences (p < 0.05). Multivariate analysis showed that the abundance and diversity of plant dwelling insects were similar. The data on insect species population densities were subjected to principal component analysis (PCA), which did not distinguish among the three varieties, Bt-9 and the non-GM, reference cultivars, during the cultivation years. However, the results of the PCA analysis were completely divided into four groups based on the yearly survey areas. Therefore, there was no evidence for a negative impact of Bt-9 on the above-ground insects and spiders.