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Also brings the potential risk of a huge gospel to mankind since the advent of 1983 's first transgenic crop , genetically modified crops safety issues are becoming increasingly attention . Governments have to develop policies and regulations to implement identity management system requirements for genetically modified crops and their products . The implementation of these regulations is to establish a stable and accurate detection of transgenic technology . This study in order to turn genes of maize MON810 on the basis of building a standard plasmid , the establishment of the MON810 qualitative PCR and fluorescence quantitative PCR detection method , the main findings are as follows : 1 , in order to obtain high-quality genomic DNA , this study compare more than the species extracted corn leaf tissue and genomic DNA of maize seed , and its optimized corn leaf tissue and seeds genomic DNA extraction method , and thus provide the basis for the detection of genetically modified crops in the nucleic acid . 2 , respectively, obtained by Tail-PCR and I - PCR method MON810 left border sequence , the degree of difficulty of the comparison of the two methods , Tail- PCR experiment is simple but the results of analysis cumbersome ; I - PCR experiment tedious but the results simple analysis , from the situation of this experiment , I-PCR for the boundary sequence . By overlap extension PCR method MON810 sequence of the left margin mt and exogenous gene CaMV35S promoter , hsp70 , cry1Ab connection into a large fragments , purified , and then connect it to the pMD18-T vector , and finally by double digestion the method of the corn within subscript gene SS Ⅱ B connected into the same vector , form suitable for MON810 detection standard plasmid pMD - of MON810 . 4 , according to the MON810 transferred to the exogenous gene , design of a targeted , CaMV35S , HSP70 and cry1Ab the primer was , and designed MON810 strains qualitative detection system for the left border sequence , the primers and the internal standard gene , thereby establishing . 5 , dye SYBR Green Ⅰ fluorescent PCR - based quantitative detection system , using a standard plasmid established linear regression equation to calculate the genetically modified content of the unknown sample . Quantitative detection sensitivity of the system to 50 copies , can fully meet the needs of international quantitative detection of genetically modified .
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