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| Study of Conventional PCR and qRT-PCR Detection Methods for Genetically Modified Soybean (Glycine max) SHZD32-1 |
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Abstract Genetically modified (GM) soybean (Glycine Max) event SHZD32-1 was developed by Chinese scientists with independent intellectual property rights, and the event has herbicide resistance by translating glyphosate resistant gene G10-EPSPS into cultivated variety 'Zhongdou 32 (Glycine Max)'. At present, GM soybean event SHZD32-1 has entered the stage of environmental testing and has a broad application prospect. So far, there are little reports about detection methods for GM soybean event SHZD32-1. In order to protect and improve the implementation of laws and regulations for GM soybean event SHZD32-1, the specific primer pairs and probes were designed based on the flanking sequences between the soybean genome and inserted exogenous fragment of SHZD32-1. Conventional PCR and qRT-PCR detection methods had been developed in this study. Specificity detection of conventional PCR and qRT-PCR found that all mixture GM samples DNA could not obtain the positive results except containing the SHZD32-1 genomic DNA, indicating the methods were good for specificity. Conventional PCR could detect 0.1% of GM SHZD32-1 event, and limit of detection (LOD) of qRT-PCR method could reach 21 copies of GM SHZD32-1 genomic DNA. The results were obtained by using the conventional PCR and qRT-PCR method for 60 repetitions under the LOD concentration. Therefore, the two methods had high specificity, good sensitivity, and strong stability. This research established GM SHZD32-1 specific PCR detection method is an important content of molecular characteristics of the evaluation of GMO, could be used to GMO safety regulation and provide technical support in China.
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Received: 26 April 2017
Published: 14 February 2018
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NI He-Jia, CHENG Xue, WANG Jing, GAO Ze-Ping, WEI Pan-Pan, TAN Jia-Li, SUN Jing, LI Hai-Tao, GAO Ji-Guo. Isolation and Screening of High Virulent Bacillus thuringiensis Strains Against Lepidopteran Pests in Some Areas of Liaoning Province[J]. 农业生物技术学报, 2019, 27(8): 1513-1520. |
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