The construction of pMD18-HT-Soybean as a calibrator plasmid and nested PCR assay for herbicide-tolerant soybeans
A multiple-target plasmid designated as pMD18-HT-Soybean, comprising part of a junction region of genetically modified soybean events A2704-12, A5547-127, MON89788 and GTS-40-3-2, and the endogenous soybean-specific lectin gene were constructed. The limit of detection for quantification of these fou...
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| Published in | European food research & technology Vol. 238; no. 3; pp. 375 - 386 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
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Berlin/Heidelberg
Springer Berlin Heidelberg
01.03.2014
Springer Nature B.V |
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| Abstract | A multiple-target plasmid designated as pMD18-HT-Soybean, comprising part of a junction region of genetically modified soybean events A2704-12, A5547-127, MON89788 and GTS-40-3-2, and the endogenous soybean-specific lectin gene were constructed. The limit of detection for quantification of these four event-specific genes using pMD18-HT-Soybean plasmid was 20 copies. Furthermore, a nested PCR detection method was developed for the above four genetically modified soybean events. LOD value of nested PCR detection was 0.005 %. The above results demonstrated that the plasmid pMD18-HT-Soybean DNA represents a valuable alternative to genomic DNA as a calibrator for the quantification of soybean event GTS-40-3-2, MON89788, A2704-12 and A5547-127 in food and feed products. And the validated results also indicated that the developed nested PCR method can be used for identification and quantification of four genetically modified soybean events and its derivates. |
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| AbstractList | A multiple-target plasmid designated as pMD18-HT-Soybean, comprising part of a junction region of genetically modified soybean events A2704-12, A5547-127, MON89788 and GTS-40-3-2, and the endogenous soybean-specific lectin gene were constructed. The limit of detection for quantification of these four event-specific genes using pMD18-HT-Soybean plasmid was 20 copies. Furthermore, a nested PCR detection method was developed for the above four genetically modified soybean events. LOD value of nested PCR detection was 0.005 %. The above results demonstrated that the plasmid pMD18-HT-Soybean DNA represents a valuable alternative to genomic DNA as a calibrator for the quantification of soybean event GTS-40-3-2, MON89788, A2704-12 and A5547-127 in food and feed products. And the validated results also indicated that the developed nested PCR method can be used for identification and quantification of four genetically modified soybean events and its derivates. A multiple-target plasmid designated as pMD18-HT-Soybean, comprising part of a junction region of genetically modified soybean events A2704-12, A5547-127, MON89788 and GTS-40-3-2, and the endogenous soybean-specific lectin gene were constructed. The limit of detection for quantification of these four event-specific genes using pMD18-HT-Soybean plasmid was 20 copies. Furthermore, a nested PCR detection method was developed for the above four genetically modified soybean events. LOD value of nested PCR detection was 0.005 %. The above results demonstrated that the plasmid pMD18-HT-Soybean DNA represents a valuable alternative to genomic DNA as a calibrator for the quantification of soybean event GTS-40-3-2, MON89788, A2704-12 and A5547-127 in food and feed products. And the validated results also indicated that the developed nested PCR method can be used for identification and quantification of four genetically modified soybean events and its derivates. |
| Author | Wang, Aoxue Qiu, Youwen Yu, Yanbo Gao, Xuejun Zhang, Minghui |
| Author_xml | – sequence: 1 givenname: Youwen surname: Qiu fullname: Qiu, Youwen organization: Detecting Center of Agricultural Genetically Modified Organism, Research Center of Life Science and Biotechnique, Northeast Agricultural University, College of Life Science, Northeast Agricultural University – sequence: 2 givenname: Minghui surname: Zhang fullname: Zhang, Minghui organization: Detecting Center of Agricultural Genetically Modified Organism, Research Center of Life Science and Biotechnique, Northeast Agricultural University, College of Life Science, Northeast Agricultural University – sequence: 3 givenname: Yanbo surname: Yu fullname: Yu, Yanbo organization: Detecting Center of Agricultural Genetically Modified Organism, Research Center of Life Science and Biotechnique, Northeast Agricultural University, College of Life Science, Northeast Agricultural University – sequence: 4 givenname: Aoxue surname: Wang fullname: Wang, Aoxue organization: College of Life Science, Northeast Agricultural University – sequence: 5 givenname: Xuejun surname: Gao fullname: Gao, Xuejun email: gaoxj5390@sina.com organization: Detecting Center of Agricultural Genetically Modified Organism, Research Center of Life Science and Biotechnique, Northeast Agricultural University, College of Life Science, Northeast Agricultural University |
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| Keywords | LOD Nested PCR Calibrator plasmid DNA Herbicide-tolerant soybean Quantitative PCR |
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| Snippet | A multiple-target plasmid designated as pMD18-HT-Soybean, comprising part of a junction region of genetically modified soybean events A2704-12, A5547-127,... |
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| SubjectTerms | Agriculture Analytical Chemistry Biotechnology Chemistry Chemistry and Materials Science Corn Deoxyribonucleic acid DNA Food Food Science Forestry Genes Genetic engineering Genetically altered foods Herbicides Laboratories Lectins Life sciences Original Paper Plant resistance Plasmids Polymerase chain reaction Soybeans Studies |
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| Title | The construction of pMD18-HT-Soybean as a calibrator plasmid and nested PCR assay for herbicide-tolerant soybeans |
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