• Food Science and Biotechnology
• /
• v.18 no.3
• /
• pp.694-699
• /
• 2009

A multiplex polymerase chain reaction (PCR) method was developed for the detection of 4 events of genetically modified (GM) soybean. The event-specific primers were designed from 4 events of GM soybean (RRS, A2704-12, DP356043-5, and MON89788). The lectin was used as an endogenous reference gene of soybean in the PCR detection. The primer pair YjLec-4-F/R producing 100 bp amplicon was used to amplify the lectin gene and no amplified product was observed in any of the 9 different plants used as templates. This multiplex PCR method allowed for the detection of event-specific targets in a genomic DNA mixture of up to 1% GM soybean mixture containing RRS, A2704-12, DP356043-5, and MON89788. In this study, 20 soybean products obtained from commercial food markets were analyzed by the multiplex PCR. As a result, 6 samples contained RRS. These results indicate that this multiplex PCR method could be a useful tool for monitoring GM soybean.

• Korean journal of applied entomology
• /
• v.54 no.4
• /
• pp.335-343
• /
• 2015

One of the primary concerns about the environmental risks of genetically modified (GM) crops is that they may have adverse effects on the local arthropod communities. In this study, we investigated whether the arthropod diversity and community structure in fields of GM rice tolerant to protoporphyrinogen oxidase (PPO)-inhibiting herbicides differ from those in non-GM (control) rice fields. The aim of this study was to assess the potential adverse effects of GM rice on the local arthropod communities. During the growing seasons in the study period, we collected arthropods from both fields by using yellow sticky traps and compared the diversity and community structure of arthropods from the two sites. Overall, the GM rice had no significant effect on the diversity of the local arthropod communities. In addition, multivariate analyses (permutational multivariate analysis of variance and nonmetric multidimensional scaling) showed that the structures of arthropod communities were not affected by the rice genotype (GM vs. non-GM), although these comparisons were made using data obtained at different sampling dates.

• Journal of the Korean Society of Food Culture
• /
• v.26 no.4
• /
• pp.331-343
• /
• 2011

A survey of consumer awareness and attitudes was conducted about genetically modified (GM) foods and the labeling regulations. The questionnaires were distributed to 4,620 consumers who lived in a variety of areas in Korea, and 4,076 people responded. The consumers were asked about knowledge, labeling information, and the source of obtaining information about GM foods. More than 11.5% of the consumers had never heard about GM foods and 86.9% of consumers had less than a normal level of knowledge about GM foods. No statistically significant relationship was found between genders, but the teachers group had moderate knowledge (p<0.001). In total, 28.4% of consumers did not know the GMO labeling regulations. They answered that the reason to buy GM food was do not know>nothing wrong>create benefit>think as safe>inexpensive. The answers to the question of what was the first benefit were: solve food shortage>functional and nutritious food>cultivate in bad condition>nothing>various cultivars. They answered that the worst factor was the next generation effect>environmental disruption. Regarding the development of GM food in Korea, males answered do not know>stronglyrecommend>defer>strongly suppress. Female answered: don't know>defer>strongly recommend>strongly suppress. More than half of the respondents did not have much information about GM foods; 88.3% of respondents answered they did not have educational experience about GM food.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.4
• /
• pp.681-684
• /
• 2007

An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

• YAKHAK HOEJI
• /
• v.45 no.3
• /
• pp.293-301
• /
• 2001

Genetically modified organism (GMO) using recombinant DNA technique has been exponentially increased, however there are still arguments for the safety of GM foods. The objective of this research was to compare the allergens of GM soybean(Roundup Ready$^{TM}$) with conventional soybeans. Each soybean extracts were prepared as crude extracts, heated extracts, and as heated and simulated gastric quid (SGF)-digested samples to characterize the stability of allergens to physicochemical treatment. Positive sera from 20 soybean-sensitive patients and control sera from 5 normal subjects were used to identify the endogenous allergens in soybeans. Specific-IgE binding activities to each soybean preparations were evaluated by ELISA and immunoblot technique. In ELISA result, IgE binding activities of positive sera to soy crude extracts generally showed two fold higher mean value than those of control sera, how-ever there was no significant difference between GM soybean and natural soybean varieties. Extracted proteins form each of the soybean preparations were separated with SDS-PAGE. The band pattern of GM soybean was very similar to those of natural soybean varieties. Immunoblots for the different soybeans revealed no differences in IgE-binding protein patterns, moreover, disclosed five prominent IgE-binding bands (75, 70, 50, 44 and 34 kDa) in crude extracts, four (75, 70, 44 and 34 kDa) in heated preparations, one (50 kDa) in heated and SGF-digested preparations. These IgE binding bands were consistent with previously reported results on the soybean. These results indicate that GM soybean (Roundup Ready$^{TM}$) is no different from natural soybean in terms of its allergen.gen.

• Korean Journal of Food Science and Technology
• /
• v.36 no.5
• /
• pp.828-832
• /
• 2004

Enzyme-linked immune sorbent assay (ELISA) was applied to quantify soy protein in fresh soybean curd (bean curd) produced by combination of genetically modified (GM) and genetically not modified (non-GM) soybeans. Antibodies against 113 and 24 kDa proteins, which appeared only in non-GM bean curd (specific band), and in both non-GM and GM bean curds (non-specific band) based on SDS-PAGE results, were prepared by immunization to rabbit. Through ELISA using either antibody, GM bean curd protein content was determined at dilution rates of $10^{-1}-10^{-6}$. Standard curve showing relationship between ELISA optical density and non-GM protein content was produced using antibody against 113 kDa protein at protein dilution between $10^{-7}\;to\;10^{-6}$, highly antigen content-dependent dilution. Bean curd prepared by random combinations of GM and non-GM soybeans were analyzed by ELISA, and standard curve was produced. Results reveal non-GM protein content of bean curd could be quantified with higher than 93% accuracy.

• Journal of Plant Biotechnology
• /
• v.47 no.4
• /
• pp.309-315
• /
• 2020

Advances in biotechnology have led to progress in crop genetic engineering to improve agricultural productivity. The use of genetically modified (GM) crops has increased, as have consumers' and regulators' concerns about the safety of GM crops to human health, and ecological biodiversity. As such, the identification of GM crops is a critical issue for developers and distributors, and their labeling is mandatory. Multiplex polymerase chain reaction (PCR) has been developed and its use validated for the detection and identification of GM crops in quarantine. Herein, we established a simultaneous detection method to identify four GM maize events. Event-specific primers were designed between the junction region of transgene and genome of four GM maize lines, namely 5307, DAS-40278-9, MON87460, and MON87427. To verify the efficiency and accuracy of the multiplex PCR we used specificity analysis, limit of detection evaluation, and mixed certified reference materials identification. The multiplex PCR method was applied to analyze 29 living, modified maize volunteers collected in South Korea in 2018 and 2019. We performed multiplex PCR analysis to identify events and confirmed the result by simplex PCR using each event-specific primer. As a result, rather than detecting each event individually, the simultaneous detection PCR method enabled the rapid analysis of 29 GM maize volunteers. Thus, the novel multiplex PCR method is applicable for living modified organism volunteer identification.

• Food Science and Biotechnology
• /
• v.15 no.1
• /
• pp.148-151
• /
• 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.

• Food Science and Preservation
• /
• v.14 no.6
• /
• pp.688-694
• /
• 2007

To evaluate the human risk of long-term intake of genetically modified (GM) rice, we carried out RT-PCR of housekeeping genes. Housekeeping genes, which show highly uniform expression in living organisms during various stages of development and under different environmental conditions, were normalized by RT-PCR. We assessed the expression of 10 common housekeeping genes (18s rRNA, 25S rRNA, UBC, UBQ5, UBQ10, ACT11, GAPDH, eEF-$1{\alpha}$, ${\beta}$-TUB, GAPDH, ${\beta}$-actin, B2m, G6pd2, Gyk, Gus, Hprt, Cyclophlin A, Tfrc, ${\alpha}$-tubulin and RPL13A) in the liver, stomach, small intestine, large intestine, kidney and spleen of mice fed GM or non-GM rice. We found no significant differences in the expression of housekeeping genes between the two groups of mice.

• Korean Journal of Agricultural Science
• /
• v.49 no.4
• /
• pp.795-807
• /
• 2022

The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10^-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD_95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.