• Korean Journal of Food Science and Technology
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• v.36 no.4
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• pp.677-682
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• 2004

Debates on safety of genetically modified (GM) crops have led to mandatory-labeling legislation of GM foods in many countries including Korea. Effects of heat, pressure, and acid treatments on degradation of DNAs or proteins in GM soybean at levels below detection limits of qualitative PCR and lateral flow strip test (LFST) methods were examined. Results showed that genomic DNAs and proteins were degraded into fragment sizes no longer possible for detection of inserted gene depending on thermal, or thermal and pressure treatment period. Detectaability of LFST for toasted meal increased in weakly treated soybean. DNA and protein detection methods were barely effective for detection of GM ingredient after $121^{\circ}C$ and 1.5 atmospheric treatment for 20 min. These results will be useful in determining GM labeling requirements of processed foods.

• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.115-123
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• 2009

In recent years, several genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by various agricultural biotechnological companies. Commercialization of GM crops will be required the assesment of risks associated with the release of GM crops. In advance of the commercial release of GM crops, developer should submit the several information on GM crops for approval. In this study, we carried out to provide the molecular data for the risk assessment of GM rice containing insect-resistant gene, modified Cry1Ac (CryIAc1). Through the molecular analysis with CryIAc1 induced GM rice, we confirmed the steady integration and expression of transgene, the transgene copy number, the adjacent region sequences of inserted gene into rice genome, and the transgene stability in progenies. For the qualitative PCR detection methods, specific primer pairs were designed on the basis of integration sequences, and construct- and event-specific detection markers were developed for leaf folder-resistant rice, Cr7-1 line. From these results, we demonstrated that the molecular data and the PCR detection methods of leaf folderresistant GM rice could be acceptable to conduct the biosafety and environment risk assessment.

• 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.

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

• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.171-182
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• 2018

Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.

• Food Science and Biotechnology
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• v.17 no.4
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• pp.829-832
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• 2008

A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect 4 varieties of genetically modified (GM) cotton. The event-specific primers were used to distinguish the 4 varieties of GM cotton (MON1445, MON15985, MON88913, and LLcotton25) using multiplex PCR. The acyl carrier protein 1 (Acp1) gene was used as an endogenous reference gene of cotton in the PCR detection. The primer pair Acp1-AF/AR containing a 99 bp amplicon was used to amplify the Acp1 gene and no amplified product was observed in any of the 13 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 cotton containing MON1445, MON15985, MON88913, and LLcotton25.

• 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.

• Korean Journal of Food Science and Technology
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• v.45 no.2
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• pp.156-160
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• 2013

A duplex polymerase chain reaction (PCR) method was developed to detect unapproved genetically modified (GM) potato (EH92-527-1) in Korea. The UDP-glucose pyrophosphorylase (UGP) gene was selected as an endogenous reference gene for potato and used to validate the specificity for 14 different crops. The primer pair EH92-F/R was designed to amplify the junction sequence between the genome and transgenic region introduced in GM potato. Its specificity was also validated using several different GM events. The detection limit of the duplex PCR method is approximately 0.05%. This duplex PCR method could be useful for monitoring cultivation of unauthorized GM potato in Korea.

• Analytical Science and Technology
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• v.28 no.2
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• pp.117-124
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• 2015

Genetically modified (GM) papaya line 55-1, which is resistant to PRSV infection, has been marketed globally. Prompt and sensitive protocols for specific detections are essential for the traceability of this line. Here, an event- and construct-specific real-time polymerase chain reaction (RT-PCR) method was established to detect 55-1. Qualitative detection was possible for fresh papaya fruit up to dilutions of 0.005% and 0.01% for the homozygous SunUp and heterozygous Rainbow cultivars, respectively, in non-GM papaya. The method was applied in the qualitative detection of 55-1 in eight types of commercially processed papaya products. Additionally, papaya products were monitored to distinguish GM papaya using the P35S and T-nos RT-PCR detection methods. As expected, detection capacity was improved via modified sample preparation and the established RT-PCR detection method. Taking these results together, it can be suggested that a suitable method for the extraction and purification of DNA from processed papaya products was established for the detection of GM papaya.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.569-572
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• 2008

A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect 3 events of genetically modified (GM) maize. The event-specific primers were used to discriminate the following 3 events of GM maize (DAS-59122-7, TC6275, and MIR604) using multiplex PCR method. The zein gene was used as an endogenous maize reference gene in the multiplex PCR detection. The primer pair Zein-FIR producing a 99 bp amplicon was used to amplify the zein gene. The primer JI-Das-F1/R1 for DAS-59122-7, JI-TC6275-F3/R3 for TC6275, and JI-MIR F1/R1 for MIR604 yielded an amplicon of 130, 162, and 197 bp, respectively. The detection limit of multiplex PCR was 1% for DAS-59122-7, TC6275, and MIR604 for one reaction.