• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.390-394
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• 2004

The genetically modified glyphosate-tolerant soybean contains the following introduced DNA sequences: the EPSPS (5-enol-pyruvylshikimate-3-phosphate synthase) gene from Agrobacterium sp. strain CP4, the 35S promoter from the cauliflower mosaic virus, and the NOS terminator from Agrobacterium tumefaciens. In the present study, detection of these introduced DNAs was performed by amplification using the polymerase chain reaction (PCR). A multiplex PCR method was also applied to prevent false positive results. When primers for 35S promoter, nos3', CTP(chloroplast transit peptide), and CP4 EPSPS (EPSPS from Agrobacterium sp. CP4) were used, positive results were obtained in PCR reactions using DNA from genetically modified glyphosate-tolerant soybeans. There were no false positive results when using DNA from non-genetically modified soybeans. The CP4 EPSPS gene was detected when less than 125 pg glyphosate-tolerant soybean DNA was amplified. Lectin Lel and psb A were amplified from both non-genetically modified and genetically modified glyphosate-tolerant soybean DNA. Multiplex PCR was performed using different primer sets for actin Sacl, 35S promoter and CP4 EPSPS. The actin gene was detectable in both non-genetically modified and glyphosate-tolerant soybeans as a constant endogenous gene. Target DNAs for the 35S promoter, and CP4 EPSPS were detected in samples containing 0.01-0.1% glyphosate-tolerant soybean, although there were variations depending on primers by multiplex PCR. Soybean seeds from five plants of non-genetically modified soybean were co-cultivated for six months with those of genetically modified soybean, and they were analyzed by PCR. As a result, they were not positive for 35S promoter, nos3' or CP4 EPSPS. Therefore, these results suggest there was no natural crossing of genes between glyphosate-tolerant and non-genetically modified soybean during co-cultivation, which indicates that gene transfer between these plants is unlikely to occur in nature.

• 한국식품과학회지
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• 제33권5호
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• pp.521-524
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• 2001

본 연구는 유전자재조합 기술에 의해 개발된 glyphosate에 내성을 가지고 있는 콩(GTS)의 모니터링을 위하여 PCR을 이용한 검출 방법에 대한 실험을 수행하였다. Glyphosate에 내성이 있는 콩에 삽입된 유전자와 표준대조 유전자인 lectin과 ferritin 유전자를 근거로 제작된 primer와 CTAB 방법으로 추출된 콩의 DNA를 template로 이용하여 PCR을 수행하였다. GTS의 검출을 위한 제작된 primer들은 GTS와 특이적으로 반응하여 증폭된 PCR 산물을 생성하였으나, non-GTS와는 PCR 산물을 생성하지 못했다. 증폭된 염기서열 분석을 통하여 GTS에 특이적인 것을확인하였으며, 약 0.05%가 포함되어 있는 GTS까지 검출이 가능함을 보였다.

• 한국식품영양과학회지
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• 제35권4호
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• pp.470-475
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• 2006

국내산 콩 품종 3종과 미국산 콩 2종(non-GMO 및 GMO glyphosate-tolerant HS2906)에 대하여 단백질함량과 분포의 차이를 아미노산 분석, 총 질소량, PAGE/densitometry법에 의 해 분석하였다. 아미노산 구성과 총 질소량에서 품종간 유의적인 차이는 없었다. 한편 단백질 추출에서 SDS/buffer 추출법이 더 효과적이었다. 총 단백질함량(380.2-423.9 mg/g)과 단백질 분포는 품종간 유사하였으나. 상세 PACE(12.5% normal gel) 결과 재래종 콩(WS82)에 비해 제초제 저항성 GMO콩(HS2906)에서 beta conglycinin(55kDa) 양이 낮게 나타났으며, 25 kDa 단백질의 경우 미국산 콩에서는 관찰되지 못한 반면에 국내산 콩 품종에서는 뚜렷하게 관찰되었으며, ${\beta}$-conglycinin은 미국산 제초제 저항성 GMO콩과 국내산 황금콩에서 상대적으로 낮은 것으로 관찰되었다. 이와 같은 결과는 normal PAGE/densitometry 분석법은 콩시료 분석에 유용하게 활용될 수 있음을 보여주었다.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.561-572
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• 2017

The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.

• 동아시아식생활학회지
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• 제15권1호
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• pp.71-77
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• 2005

Proximate analysis, mineral and fatty acid composition of three conventional domestic soybean cultivars and two imported ones including glyphosate-tolerant HS2906 were evaluated by AOAC method, ICP-AES and gas chromatography. There were several differences in the proximate analysis among three conventional domestic soybean cultivars ; higher crude fat in the cultivar Hwanggumkong, higher crude protein in Pungsankong, and higher carbohydrate and crude ash in Duyukong. The ranges of contents of proximate components of domestic cultivars were similar to the data previously reported. There were no significant differences in proximate analysis between conventional soybean WS82 and glyphosate-tolerant HS2906 ; 23.55-23.90% of crude fat, 34.22-35.55% of crude protein, 6.25-6.45% of crude ash, and 25.35-26.47% of carbohydrate. The mineral and fatty acid compositions of HS2906 were similar to those of conventional soybeans previously reported.

• 한국식품과학회지
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• 제36권3호
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• pp.369-374
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• 2004

유전자재조합 콩 Roundup Ready는 Agrobacterium sp. CP4에서 유래한 유전자를 함유하고 있으며 이 삽입 유전자에서 5-enolpyruvylshikimate-3-phosphate synthase(CP4EPSPS)가 발현된다. 본 연구실에서는 이 단백질을 유전자재조합 콩과 시중에서 구입한 두부 시료에서 CP4EPSPS 특이 단클론 및 다클론 항체를 이용하여 immunoblotting법으로 검사하였다. 분리 정제된 CP4EPSPS 재조합 단백질은 단클론 또는 다클론 항체를 이용할 때 각각 $0.006{\mu}g$ 또는 $0.0006{\mu}g$의 검증 한계치로 확인할 수 있었다. 유전자재조합 콩에 발현된 CP4EPSPS 검증 한계치는 단클론 또는 다클론 항체를 이용할 때 각각 $0.001{\mu}g$과 $0.0001{\mu}g$이었다. 다클론 항체를 이용하여 조사된 9종의 두부에서는 2종에서 양성결과를 확인하였다. 본 연구에서 생산된 단클론 및 다클론 항체를 이용하여 확립된 immunoblotting법은 콩 가공식품 중의 glyphosate 내성 유전자재조합 콩 검사에 적용될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.25-31
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• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• Food Science and Biotechnology
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• 제15권1호
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• pp.91-95
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• 2006

Effects of cultivars, cooking, and processing on hemagglutinin activity were evaluated by observing macroscopic hemagglutination using serial twofold dilution of trypsinized human blood type-O or rabbit blood. Hemagglutinin activity was expressed as maximal geometric dilution fold. Agglutination of rabbit blood was more sensitive compared to human blood. Hemagglutinin activities of glyphosate-tolerant soybean, HS2906, and imported conventional soybeans were not statistically different, although significant differences were observed among conventional soybean cultivars cultivated in Korea (286 to 1535 HU/mg protein). Time required to reach fifty percent inhibition of hemagglutinin activity ($IT_{50}$) value decreased with increasing cooking temperature and pressure. Most effective conventional cooking method to inhibit hemagglutinin activity was pressure-cooking ($IT_{50}$: 1.36 min). Calculated activation energy based on reaction rate constant was 4.88 kcal. No hemagglutinin activities were detected in processed soybean products such as tofu, soybean paste, and soysauce.

• Preventive Nutrition and Food Science
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• 제10권1호
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• pp.6-10
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• 2005

The trypsin inhibitor activity (TIA) of various soybean cultivars was evaluated by measuring the inhibition of trypsin activity using N-benzoyl-DL-arginine-p-nitro-anilide (BAPNA) as the substrate. The TIA values of eleven white shelled soybean cultivars including a glyphosate-tolerant soybean (16.58 to 17.90㎎/g) were not significantly different among cultivars. Black shelled soybeans had higher TIA values, ranging from 40.09 to 52.11㎎/g, compared to white shelled soybeans (p<0.05). When the TIA of commercially processed soybean foods were determined, no TIA was detected in soysauce, tofu and soybean paste. During conventional moist heating, the IT/sub 50/ (Time required to reach 50% inhibition of TIA) values were decreased as heating temperature and cooking pressure increased. The IT/sub 50/ values of moist heating were estimated to be 91.68, 37.71 and 19.50 min at 60, 80 and 100℃, respectively. The IT/sub 50/ value of microwave cooking was 4.75 min at medium heat, while that of the pressure cooking at 120℃ was only 2.62min. Moreover, there was a negative relationship between temperature and IT/sub 50/ values (R=0.92, p<0.01). The TIA of soybean sprouts was completely inactivated after heating at 100℃ for 5 min, although fresh soybean sprouts showed one fifth of the TIA value of white shelled soybeans. Based on our results, pressure cooking is the most effective cooking method to reduce TIA in soybeans.