| To ensure the labeling requirements of GM food in China, seven GM crops,namely GM maize MIR162, BVLA430101, MON88017, GM soybean MON89788,GTS40-3-2and GM canola MS1, RF1, were selected as research object. The purposeof this study was to establish the event-specific detection method of these seven linesbased on oligonucleotide microarray.Four aspects that related to GMOs identification are covered in this study.In first chapter, we reviewed the implementation of genetic engineering inagricultural and food industry and compared the labelling systems between differentcountries, and then introduced the detection methods of transgenic products. We alsointroduced the backgrounds of MIR162, BVLA430101, MON88017, MON89788,GTS40-3-2, MS1and RF1, briefly.2The primers were designed based upon the revealed flanking sequences ofMIR162, BVLA430101, MON88017, MON89788, GTS40-3-2, MS1and RF1. Thequalitative PCR detection methods were of high specificity and sensitivity (0.1%).After validated by six external laboratories, results showed that the qualitative PCRdetection method of MIR162could specifically detect the samples of MIR162withthe detection sensitivity about0.1%, a good repeatability and reproducibility, whichsuggested that the developed qualitative PCR methods can be used for theidentification of GM maize MIR162as national standard.3Based on the unique and specific PCR sequences, a40bp event-specific Oligoprobes with a high specificity have being developed. Then the transgenic detectionDNA chips were established. The results indicated that the probe which designed withverification of specificity and sensitivity could recognize PCR products with strongerspecificity and higher sensibility (0.01%). 4One set of3×multiplex PCR system and two sets of4×multiplex PCR systemsfor detecting and identifying transgenic crops were set up and optimized. Differentevents could be detected in one oligonucleotide microarray, which increased thedetection throughput. |
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