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| Construction and Application of Standard Plasmid Molecules for Screening Detection of Four Major Genetically Modified Crops |
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Abstract Plasmid molecule worked as reference material has been shown to be a good alternative in genetically modified organisms (GMOs) detection, and can promptly satisfy the regulatory requirements of GMOs. In this study, according to the transgenic information of 4 main crops including rice (Oryza sativa L.), maize (Zea mays L.), soybean (Glycine max) and rape (Brassica napus), 4 types of plasmids for GMO screening were developed and confirmed. The constructed pMD-rice contained 7 target elements including promoter of Cauliflower mosaic virus (CaMV35S), terminator of nopaline synthase (tNOS) and selective marker genes, such as biolaphos resistance (Bar), neomycin phosphotransferase (NPTⅡ) and hygromycin B phosphotransferase (Hpt), insect resistant gene of Bacillus thuringiensis (Bt) and a rice endogenous reference gene of sucrose phosphate synthase (SPS) were selected. The constructed pMD-maize was consisted of 4 target sequences including tNOS, CaMV35S, Bar and zSSⅡb as endogenous reference genes. The constructed pMD-soybean contained 5 target elements including CaMV35S, tNOS and 2 exogenous genes cowpea trypsin inhibitor gene (CpTi) and enolpyruvylshikimate phosphate synthase (EPSPS), and the Lectin precursor protein gene as endogenous reference gene. The constructed pMD-rape contained 8 target elements including CaMV35S, Figwort mosaicvirus 35S (FMV35S), nopaline synthase promoter (P-nos) and tobacco tapetum specific promoter (PTa29), tNOS, terminator-35S (T-35S) and the 7th gene of TL-DNA (g7) and high mobile group protein gene (HMG) as endogenous reference genes. In theory, all elements of the 4 plasmids covered 90% of the approved transgenic events of 4 major crops as maize, rape, soybean and rice in the world, and they covered 100% of transgenic events that had been approved in China. Then the constructed plasmids and reference materials were comparatively analyzed and all the results demonstrated that the constructed plasmids were suitable for GMO screening detection.
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Received: 23 March 2015
Published: 09 July 2015
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