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Detection of recombinant DNA of genetically modified (GM) soybeans in heat-treated GM soybeans and commercial natto
Affiliation:1. Institute of Agrophysics, Polish Academy of Sciences, Doświadczalna 4, 20-290 Lublin, Poland;2. Maria Curie-Skłodowska University, Faculty of Chemistry, Maria Curie-Skłodowska Sq. 3, 20-031 Lublin, Poland;1. The Second Hospital of Dalian Medical University, Dalian 116023, People''s Republic of China;2. Hands and Feet Microsurgery, The Second Hospital of Dalian Medical University, Dalian 116023, People''s Republic of China;3. Orthopedic Department, Changchun University of Traditional Chinese Medicine Affiliated Hospital, Changchun 130021, People''s Republic of China;1. Neuropsychiatry Group, Neurology Division, University Hospital, Universidade Federal de Minas Gerais, 30130-100 Belo Horizonte, MG., Brazil;2. Infectious Diseases Reference Center-CTR-DIP Orestes Diniz, Municipal Health Division/Universidade Federal de Minas Gerais, 30150-260 Belo Horizonte, MG., Brazil;3. Tropical Medicine Program, School of Medicine, Universidade Federal de Minas Gerais, Belo 30130-100 Horizonte, MG., Brazil
Abstract:We examined the detection of recombinant DNA of genetically modified (GM) soybeans in heat-treated GM soybeans and commercial natto. Genomic DNA was extracted from heat-treated GM soybeans and natto using the cetyltrimethylammonium bromide (CTAB) or alkaline lysis methods. First, primer pairs amplifying the junction region between CTP and CP4EPSPS in recombinant soybean were designed; they gave PCR products of band sizes, 100, 110, 120, 130, 140, and 150 bp. When DNA solution extracted from heat-treated GM soybeans by the alkaline lysis method was applied to polymerase chain reaction (PCR), PCR products of the expected 100, 110, 120, 130, 140, and 150 bp were detected by agarose gel electrophoresis. However, PCR products were not detected in DNA extracted from heat-treated GM soybeans by the CTAB method, and no PCR products were detected in either extract from natto. Next, PCR using primer pairs amplifying the junction region between NOS and part of CP4EPSPS were designed; they gave PCR products of 100, 110, 120, 130, 140, and 150 bp. The expected PCR products from all DNA extracts were detected (heat-treated GM soybeans by CTAB, 100–140 bp; heat-treated GM soybeans by alkaline lysis, 100–150 bp; natto by CTAB, 100–130 bp; and natto by alkaline lysis, 100–150 bp). These results indicate that judicious selection of DNA extraction methods and target sequences is important to detect DNA from natto and recombinant DNA can be detected in natto.
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