Establishment of a genetically modified crops screening system based on universal multiplex-PCR

• Main Authors: I-Jen Lu, 盧奕蓁
• Other Authors: Tzu-Ming Pan
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/21750554524561327599

碩士 === 國立臺灣大學 === 微生物與生化學研究所 === 97 === The PCR targets of this multiplex PCR detection system include six transgenic elements: cauliflower mosaic virus (CaMV) 35S promoter, Agrobacterium tumefaciens nopaline synthase (nos) promoter, Agrobacterium tumefaciens nopaline synthase (nos) terminator, neomycin phosphotransferase II (npt2) gene, 5-enolpyruvylshikimate-3-phosphate synthase (CP4 epsps) gene and phosphinothricin N-acetyltransferase (pat) gene. It can screen out most of the commercialized GM events. To calculate the classifying efficiency, we use the statistical software (SPSS ver. 12.0) to simulate the classification of all GM events according to the expected patterns of the multiplex PCR results. The classification exhibits that the biggest group of the 24 positive groups only contains 12 GM events. Therefore to identify the exact event of unknown sample by this system combined with event-specific PCR detection methods, only have to run 12+1 polymerase chain reactions. Moreover, the primer pairs of this system proved to have specificity to their targets and coamplify every target in one PCR reaction.