• 한국식품위생안전성학회지
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• 제33권3호
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• pp.193-199
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• 2018

본 연구에서는 유전자 변형 옥수수(GM 옥수수)에 특이한 단크론성 항체를 개발하고 이에 대한 특성을 확인하는 연구를 수행하고자 하였다. 먼저 형질전환 대장균으로부터 PAT 단백질을 발현시킬 수 있는 시스템을 확립하였고, 재조합 PAT 단백질을 대량 생산하여 항원으로 사용하였다. 준비된 항원을 면역한 결과 재조합 PAT 단백질의 항원성은 매우 높은 것으로 확인되었으며, 세포융합과 클로닝을 통해 12 종의 hybridoma를 확립하였고 western blot 결과 10 종의 hybridoma가 재조합 PAT 단백질과 강한 반응성을 나타내었다. 10종의 hybridoma가 생산하는 항체가 실제 GM 옥수수에 반응하는지를 추가의 western blot으로 분석한 결과 2종의 단크론성 항체(PATmAb-7 and PATmAb-12)가 재조합 PAT 단백질뿐만 아니라 실제 GM 옥수수 중 PAT와 반응하는 것으로 확인되었다. 항체를 대량 생산하고 정제한 후 2종의 항체는 SDS-PAGE 상에서 대표적인 항체의 분리패턴(heavy와 light chain)을 나타내었고, 전형적인 $IgG_1$과 ${\kappa}$ type으로 확인되었다. 정제된 단크론성 항체는 특성을 조사한 결과 다른 GMO에서 발현될 수 있는 재조합 단백질과 non-GM 옥수수 추출물에는 반응성이 없고 PAT 단백질에만 특이적으로 반응하는 것을 확인할 수 있었다. PATmAb-7 를 이용한 간접효소면역분석법의 검출한계는 0.3 ng/mL 수준으로 기준의 유전자변형 콩 면역분석법과 비교했을 때 높은 민감도를 나타내었다. 이상의 결과로 볼 때 개발된 2종의 항체(PATmAb-7 and PATmAb-12)는 GM 옥수수에서 발현되는 PAT 단백질에 특이적으로 반응하는 항체로 확인되었고, 2종의 항체를 이용한 면역분석법과 바이오센서의 개발 가능성을 제시할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권5호
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• pp.764-772
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• 2003

Two genetic constructs used to confer improved agronomic characteristics, namely herbicide tolerance (HT) in maize and soyabean and insect resistance (Bt) in maize, are considered in respect of feeding to farm livestock, animal performance and the nutritional value and safety of animal products. A review of nucleic acid (DNA) and protein digestion in farm livestock concludes that the frequency of intact transgenic DNA and proteins of GM and non-GM crops being absorbed is minimal/non existent, although there is some evidence of the presence of short fragments of rubisco DNA of non-GM soya in animal tissues. It has been established that feed processing (especially heat) prior to feeding causes significant disruption of plant DNA. Studies with ruminant and non-ruminant farm livestock offered GM feeds demonstrated that animal performance and product composition are unaffected and that there is no evidence of transgenic DNA or proteins of current GM in the products of animals consuming such feeds. On this evidence, current HT and Bt constructs represent no threat to the health of animals, or humans consuming the products of such animals. However as new GM constructs become available it will be necessary to subject these to rigorous evaluation.

• Food Science and Biotechnology
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• 제17권3호
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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.

• Preventive Nutrition and Food Science
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• 제17권4호
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• pp.274-279
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• 2012

For the quantitative analysis of genetically modified (GM) maize in processed foods, primer sets and probes based on the 35S promoter (p35S), nopaline synthase terminator (tNOS), p35S-hsp70 intron, and zSSIIb gene encoding starch synthase II for intrinsic control were designed. Polymerase chain reaction (PCR) products (80~101 bp) were specifically amplified and the primer sets targeting the smaller regions (80 or 81 bp) were more sensitive than those targeting the larger regions (94 or 101 bp). Particularly, the primer set 35F1-R1 for p35S targeting 81 bp of sequence was even more sensitive than that targeting 101 bp of sequence by a 3-log scale. The target DNA fragments were also specifically amplified from all GM labeled food samples except for one item we tested when 35F1-R1 primer set was applied. A reference plasmid pGMmaize (3 kb) including the smaller PCR products for p35S, tNOS, p35S-hsp70 intron, and the zSSIIb gene was constructed for real-time PCR (RT-PCR). The linearity of standard curves was confirmed by using diluents ranging from $2{\times}10^1{\sim}10^5$ copies of pGMmaize and the $R^2$ values ranged from 0.999~1.000. In the RT-PCR, the detection limit using the novel primer/probe sets was 5 pg of genomic DNA from MON810 line indicating that the primer sets targeting the smaller regions (80 or 81 bp) could be used for highly sensitive detection of foreign DNA fragments from GM maize in processed foods.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1273-1278
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• 2009

This study aimed to evaluate the potential allergenicity of Cry proteins in insect-resistant genetically modified (GM) maizes (Bt11, MON810, and MON863) using serum screening tests. Serum samples were obtained from Korean children (0-15 years old) with allergic symptoms who had positive maize-specific IgE. The levels of serum specific IgE was measured by the Phadia ImmunoCAP system and considered as positive when they are 0.35 kU/L or higher. Cry proteins (Cry1Ab in Bt11, mCry1Ab in MON810, and Cry3Bb1 in MON863) were expressed in Escherichia coli and purified for serum screening. The reactivity of purified Cry proteins was confirmed by IgE immunoblots in 50 patients (maize-sensitized patients). There was no reaction between Cry proteins and sera from maize-sensitized patients. Our results suggest that these Cry proteins are not likely to cause allergic reactions. Further studies using more sera from patients with true clinical allergies are needed to evaluate the potential allergenicity of novel proteins in GM maize.

• Journal of Plant Biotechnology
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• 제38권2호
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• pp.143-150
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• 2011

1994년 처음으로 GM 토마토인 Flavr Savr가 시장에 나온 이후, 2010년 현재 140여 품목의 GM식물이 전 세계적으로 상업화되었다. GM식물들에 대한 안전성 승인여부의 확인 및 표시제관리를 위하여 이들 GM식물내로 도입된 삽입유전자의 정보를 이용한 검정방법이 도입되었으며, 또한 도입유전자의 발현된 단백질을 분석하기 위하여 정성 및 정량을 위한 면역학적 방법이 도입되었다. 본 총설에서는 국내 외적으로 개발된 콩, 옥수수, 카놀라, 면화 등의 GM식물에 적용된 multiplex PCR, real-time PCR 방법과 최신 개발 중인 microarray, 나노기술 등을 활용한 방법들을 조사하였다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2021년도 추계학술대회
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• pp.226-226
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• 2021

• Journal of Plant Biotechnology
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• 제36권2호
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• pp.106-114
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• 2009

Genetic components introduced into approved GM crops are a key subject for safety assessment and provide a basis for the development of detection methods for GM crops. In order to understand the genetic components in approved GM crops comprehensively, we screened the genetic vector maps of GM crops that had been approved for commercialization around the world. A total of 64 varieties from 5 major GM crop species (maize, canola, cotton, soybean, and tomato) were subjected to analysis. The genetic components included genes, promoters, terminators, and selection marker. This survey may be useful for researchers who develop GM crops and methods for detecting GM crops.

• 농업과학연구
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• 제43권2호
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• pp.215-220
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• 2016

The global cultivation area of genetically modified crops (GM crops) has been increasing every year. Cultivation of GM crops is not only beneficial to the economy but also has positive effects on the environment in decreasing the use of agrochemicals, chemical fertilizers, and agricultural machinery. However, there have been controversies about the admixture of GM crops and non-GM crops and the unintentional release of GM crops to the environment. Especially in Korea, where consumption of agricultural products is import-dependent, the economic importance of GM crops has been a significant issue. The Act on import and distribution of GM crops was established in 2001 to start the management of GM crops in Korea. Recently, the imported amount of GM crops to Korea has reached over 10 million tons and is increasing very rapidly; consequently, the potential environmental impact of GM crops is becoming a big issue in Korea. In Japan, the discovery of imported GM canola plants around ports in 2005 raised awareness of the unintentional release of GM crops. In Korea, GM maize plants were also found in port and feed factory surroundings from 2005 to 2007. It is now necessary to monitor imported GM crops by tracing distribution, transport process for practical environmental risk assessment. Possible gene transfer from GM crops to non-GM crops should also be investigated in the cultivation area and the surroundings as well.

• 생명과학회지
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• 제30권5호
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• pp.411-419
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• 2020

GM 작물의 개발은 경제적 중요성을 갖게 되었고 상업적인 GM 작물을 재배하는 국가들은 1960년대 이후 지속적으로 증가하고 있다. 비록 GM 작물의 경작과 상업화는 경제적 중요성을 얻었지만, 여전히 non-GM 작물에 비해 그들의 농업적 특성을 평가할 필요가 있다. 본 연구는 유전공학에서 사용된 방법의 결과로 발생할 수 있는 의도하지 않은 문제 발생 여부를 확인하기 위해 내건성 유전자 CaMsrB2를 포함한 GM 쌀과 non-GM 쌀의 농업적 특성을 내건성 온실과 관개수답에서 평가했다. 관개수답에서는 GM벼와 non-GM벼의 모든 농업형질에서 유의미한 차이가 없었다. 그러나 내건성온실에서 수수와 수량에서 GM벼와 non-GM벼에서 유의미한 차이가 있었다. 따라서 본 연구 결과는 CaMsrB2 유전자를 함유한 GM벼가 내건성 조건에서 non-GM 쌀에 비해 경제적 가치가 우수하다는 것을 시사한다. 이 결과는 또한 CaMsrB2 유전자를 함유한 GM 벼는 가뭄에 취약한 지역에서 안정적으로 수량을 유지 하면서 재배 가능 하다.