• Journal of Microbiology and Biotechnology
• /
• v.14 no.2
• /
• pp.390-394
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.33 no.5
• /
• pp.521-524
• /
• 2001

A method using PCR was developed for the monitoring of glyphosate tolerant soybean (GTS) produced by the DNA recombination technique. We designed 3 pairs of specific oligonucleotide primers based on the gene sequences inserted in soybean and in lectin and ferritin genes as internal standards. Template DNAs were isolated from soybeans by the modified hexadecyl trimethyl ammonium bromide (CTAB)method and used for PCR with different primer sets. PCR, used with specific primer sets for GTS detection, showed the amplified DNA fragments with GTS template DNA but no product showed with non-GTS template. PCR amplified products were confirmed by DNA sequencing and were detected for up to 0.05% of GTS template DNA.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.35 no.4
• /
• pp.470-475
• /
• 2006

In order to elucidate the differences of protein profiles among soybean cultivars, the protein composition of three conventional domestic soybean cultivars and two imported ones including glyphosate-tolerant HS2906 was analyzed by total nitrogen measurement, amino acid analysis and PAGE/densitometry. There were no statistically significant differences in the levels of any amino acid, including aromatic amino acids, between glyphosale-tolerant soybean and the conventional soybean WS82. In the extraction of protein, the SDS/buffer system was more efficient than the defatting/water system. The SDS-PAGE/densitometry analysis showed that there was a similar profile of proteins among cultivars, although the amount of total protein ranged from 380.2 mg/g to 423.9 mg/g. In addition, there was no discernable difference of protein profile between glyphosate- tolerant soybean (total protein amount, 380.2 mg/g) and the conventional soybean WS82 (390.2 mg/g), although the amount of ${\beta}$-conglycinin (55 kDa) was lower in glyphosate-tolerant soybean. Meanwhile, the amount of 25 kDa protein was greater in domestic soybean cultivars than imported ones. Thus, normal PAGE/ densitometry method would be useful to analyze the difference in protein profiles of soybean proteins, and furthermore Evaluate the protein profile of proteins between GMO and conventional soybean.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.3
• /
• pp.561-572
• /
• 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.

• Journal of the East Asian Society of Dietary Life
• /
• v.15 no.1
• /
• pp.71-77
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.36 no.3
• /
• pp.369-374
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.1
• /
• pp.25-31
• /
• 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
• /
• v.15 no.1
• /
• pp.91-95
• /
• 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
• /
• v.10 no.1
• /
• pp.6-10
• /
• 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.