• Title/Summary/Keyword: genetically modified (GM) food

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Genetic information analysis for the development of an event-specific PCR marker for herbicide tolerance LM crops

  • Do Yu, Kang;Myung Ho, Lim;Soo In, Sohn;Hyun Jung, Kang;Tae Sung, Park
    • Korean Journal of Agricultural Science
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    • v.48 no.4
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    • pp.1051-1065
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    • 2021
  • Recent times have seen sustained increases in genetically modified (GM) crops not only for cultivation but also for the utility of food and feed worldwide. Domestically, commercial planting and the accidental or unintentional release of living modified (LM) crops into the environment are not approved. Many detection methods had been devised in an effort to realize effective management of the safety of agricultural genetic resources. In order to develop event-specific polymerase chain reaction (PCR) markers for LM crops, we analyzed the genetic information of LM crops. Genetic components introduced into crops are of key importance to provide a basis for the development of detection methods for LM crops. To this end, a total of 18 varieties from four major LM crop species (maize, canola, cotton, and soybeans) were subjected to an analysis. The genetic components included introduced genes, promoters, terminators and selection markers. Thus, if proper monitoring techniques and single or multiplex PCR strategies that rely on selection markers can be established, such an accomplishment can be regarded as a feasible solution for the safe management of staple crop resources.

Multiplex PCR Detection of the GT73, MS8xRF3, and T45 Varieties of GM Canola

  • Kim, Jae-Hwan;Kim, Tae-Woon;Lee, Woo-Young;Park, Sun-Hee;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.104-109
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    • 2007
  • A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect three varieties of genetically modified (GM) canola. The construct-specific primers were used to distinguish the following three varieties of GM canola; GT73, MS8xRF3, and T45, using multiplex PCR. The FatA (fatty acyl-ACP thioesterase) gene was used as an endogenous canola reference gene in the PCR detection. The primer pair Canendo-FIR containing a 105 bp amplicon was used to amplify the FatA gene and no amplified product was observed in any of the 15 different plants used as templates. The GT73-KHUF1/R1 primer recognized the 3'-flanking region of GT73, resulting in an amplicon of 125 bp. The Barstar-F1/MS8xRF3-R primer recognized the junction region of bars tar and the NOS terminator introduced into MS8xRF3, resulting in a 162 bp amplicon, and the T45-F2/R2 primer recognized the junction region of PAT and the 35S terminator introduced into T45, resulting in an amplicon of 186 bp. This multiplex PCR allowed for the detection of construct-specific targets in a genomic DNA mixture of up to 1% GM canola containing GT73, MS8xRF3, and T45.

Detection of Genetically Modified Soybean in Tofu and Biji using PCR and Immunological Methods (PCR 방법과 면역학적 분석법을 이용한 두부와 비지에서 GM 콩의 검출법)

  • Kim, Myo-Young;Kim, Jae-Hwan;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.77-81
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    • 2005
  • To monitor GM soybean in soybean processed foods, tofu and biji, we prepared tofu and biji containing 0%, 1%, 3%, 5% and 100% GM soybean, respectively. We examined epsps gene inserted in soybean by PCR and EPSPS protein expressed in soybean using western blotting and lateral flow strip test to compare the sensitivity of these methods. A PCR product of 123 bp inserted in GM soybean was detected in all tofu and biji containing 1%, 3%, 5% and 100% GM soybean with the exception of 0% samples; however, the size of 600 bp inserted in GM soybean was only detected in tofu containing 100% soybean and in biji containing 5% and 100% soybean. In the protein level, GM soybean product was only detected in tofu and biji containing 100% GM soybean by western blotting. In addition, only biji containing 100% GM soybean was detected by lateral flow strip test. We concluded that in order to detect GM soybean efficiently in processed food, the PCR method is more sensitive than immunological methods. With the PCR method, small size product with approximately 100 bp in PCR product is sensitive to detect GM soybean in processed foods.

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.

Influence of Cooking on Nutrient Composition in Provitamin A- Biofortified Rice (가열조리가 프로비타민 A 강화 쌀의 영양성분에 미치는 영향)

  • Lee, Young-Tack;Kim, Mi-Hyun;Im, Ji-Soon;Kim, Jae-Kwang;Ha, Sun-Hwa;Lee, Si-Myung;Kweon, Soon-Jong;Suh, Seok-Chul
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.683-688
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    • 2011
  • This study was conducted to investigate the effects of different cooking methods on the nutrient composition of genetically modified (GM) provitamin A(PA)-biofortified rice. PA-biofortified rice was subjected to different cooking methods, namely boiling and a soaking, steaming and roasting process. The proximate components (starch, protein, lipid and ash) of raw PA-biofortified rice were similar to those found in the parental non-GM rice, and were not significantly affected by a conventional boiling treatment. When compared with raw rice, boiled PA-biofortified rice showed a similar nutrient composition, despite a slight reduction in a majority of its amino acid contents. However, the PA-biofortified rice that underwent the soaking/steaming/roasting process exhibited a significant reduction in a majority of its amino acids and mineral contents. This procedure also led to a significant reduction in carotenoid contents. The overall results of this study demonstrate that using a conventional boiling method for PA-biofortified rice retains nutrients better than a soaking/steaming/roasting method.

Development of glufosinate-tolerant GMO detection markers for food safety management (식품안전관리를 위한 제초제 glufosinate 특이적 GM 작물 검출마커 개발)

  • Song, Minji;Qin, Yang;Cho, Younsung;Park, TaeSung;Lim, Myung-Ho
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.40-45
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    • 2020
  • Over 500 genetically modified organisms (GMOs) have been developed since 1996, of which nearly 44% have glufosinate herbicide-tolerant traits. Identification of specific markers that can be used to identify herbicide-tolerant traits is challenging as the DNA sequences of the gene(s) of a trait are highly variable depending on the origin of the gene(s), plant species, and developers. To develop specific PCR marker(s) for the detection of the glufosinate-tolerance trait, DNA sequences of several pat or bar genes were compared and a diverse combination of PCR primer sets were examined using certified reference materials or transgenic plants. Based on both the qualitative and quantitative PCR tests, a primer set specific for pat and non-specific for bar was developed. Additionally, a set of markers that can detect both pat and bar was developed, and the quantitative PCR data indicated that the primer pairs were sensitive enough to detect 0.1% of the mixed seed content rate.

In vitro Digestibility Assessment of CP4EPSPS in GM Soybean under Different Conditions of Simulated Gastric Fluid and Preheating (인공위액조건과 예열처리에 따른 GM 콩 도입단백질(CP4EPSPS)의 소화성 평가)

  • Choi, Mi-Hee;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1310-1314
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    • 2012
  • Gastrointestinal digestibility of new proteins inserted in the food supply is a significant parameter for assessing the safety of GM foods based on the assumption that digestive stability is undesirable. In this study, we performed in vitro digestion of CP4EPSPS, a new protein, expressed in genetically modified (GM) soybean in order to evaluate its digestibility in three different ratios of simulated gastric fluid with preheating. Ratios of GM soybean to simulated gastric fluid were 2:2, 2.5:1.5, and 1.5:2 and preheating was conducted at $100^{\circ}C$ for 5 min. Electrophoresis and Western blotting were used to confirm changes in soybean protein patterns and CP4EPSPS gene expression after in vitro digestion. At ratios in which the amount of gastric fluid was equal to (2:2) or relatively higher than that of soybean (1.5:2), no CP4EPSPS (47.4 kDa) protein was detected after 15 seconds of simulated gastric fluid incubation, the earliest time interval evaluated. However, when the ratio of GM soybean to gastric fluid was 2.5:1.5, CP4EPSPS was detected in 5 min and gradually decreased according to time. After preheating, no CP4EPSPS protein was detected after 15 seconds under all conditions. From these results, we concluded that the digestibility of CP4EPSPS in simulated gastric fluid increased upon preheating. Accordingly, we suggest that it is important to account for the ratio of gastric fluid to GM food in in vitro digestibility assessment models of GM food.

Qualitative PCR Detection of vitamin E-enriched GM Perilla (비타민 E 강화 유전자변형 들깨에 대한 정성 PCR 분석법)

  • Kim, Jae-Hwan;Ahn, Ji-Hye;Song, Hee-Sung;Kim, Kyung-Hwan;Kim, Dong-Hern;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.192-195
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    • 2006
  • For the development of a qualitative PCR detection method for genetically modified perilla (Perilla frutescens), perilla species-specific gene, KAS-I (Beta-ketoacyl-ACP synthase I), was selected and validated as suitable for the use as an endogenous reference gene in perilla. Primer specificity was first tested by the means of qualitative PCR analysis. The primer pair Pfru3-F/R amplifying the perilla endogenous gene, KAS-I, gave rise to an amplicon 95 bp. No amplified product was observed when DNA samples from 15 different plants were used as templates. Qualitative PCR detection method was assayed with vitamin E-enriched GM Perilla developed in Korea. For the qualitative PCR detection method, the construct-specific detection primer pairs were constructed. The primer pair TMTO-F/R amplifying the junction region of TMT (${\gamma}$-tocopherol methyltransferase) gene and OCS (Octopine synthase) terminator introduced in GM perilla gave rise to an amplicon 148 bp.

The Study of Environmental Risk Assessment for Fluorescent Genetically Modified Silkworms (형광단백질 발현 유전자변형 누에(Bombyx mori )의 환경위해성 평가연구)

  • Kim, Hyunjung;Jung, Chuleui;Goo, Taewon;Yi, Hoonbok
    • Korean journal of applied entomology
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    • v.53 no.3
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    • pp.199-207
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    • 2014
  • It is true that the proper environmental risk assessments for many GM (Genetically Modified) insects almost have not been executed in Korea. Therefore, we tested the environmental risk assessment about GM silkworms if there is any difference between GM silkworms and non-GM silkworms by the following three measurements. First, we measured their mobility in the breeding environment conditions with food and without food. Secondly, we measured their viability at the artificial extreme environmental conditions (low and high temperature and humidity, absent/present of foods,) after escaping from their breeding environments. Thirdly, we observed the number of laying eggs and their hatchability between GM silkworms and non-GM silkworms with four different pair experiments. The mobility of GM silkworms and non-GM silkworms statistically did not differ, and the egg productivity and hatchability were not also different. The hatchability by couple of GM female silkworms and non-GM male silkworms was lower than by non-GM male and female couple between the GM silkworms and non-GM silkworms, and there was statistically different. Relatively, the viability of GM silkworms was lower than non-GM silkworms. We could not exactly test for viability of silkworms in low temperature conditions because of their hibernating. Although there was any difference in viability and hatchability between GM silkworms and non-GM silkworms, all ability of GM silkworms was lower than non-GM silkworms. Conclusively, the environmental risk of GM silkworm was relatively lower than non-GM silkworm in this study.

Changes in DNA Fragments in Bt11 Corn Caused by Processing Conditions and Their Monitoring (가공조건에 따른 GM corn Bt11의 유전자 변화와 모니터링)

  • Lee, Cheol-Su;Kim, Young-Chan;Hwang, Soon-Wook;Kang, Sang-Mo
    • Korean Journal of Food Science and Technology
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    • v.36 no.2
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    • pp.299-305
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    • 2004
  • Genetically Modified (GM) corn 'Bt11' was developed to promote insect resistance using crylA (b) gene derived from Bacillus thuringiensis. Effects of heat, pressure, and ${\alpha}-amylase$ on DNA fragment degradation in Btll corn were examined through PCR. Whereas DNA fragment degraded completely within 4 min at $150^{\circ}C$ and by autoclave, most remained after oil-frying, boiling, and drying-autoclave. Treatment of ${\alpha}-amylase$ enzyme did not affect DNA fragment degradation. Among 65 corn-processed foods analyzed, 9 were detected as GM corn-containing foods(13.6%).