| Due to we cann' t calculate accuratelly the efect on the safty of human and environment produced by transgene with current science and technology, consequently, it becomes the major measures to limit the transgenic crops enter our market in large numbers by detection and identification. The major purpose of transgenic detection is to identify whether they are GMOs or not, how many about the transgene ingredient and whether they are approved and permit to import by our government, etc. Previously, some transgenic detectin methods had been reported, but the detection method is simplistic not the general screening and identifying method. In abroad, the study of integration site used for transgenic detection had just begun.In this study, according to the collection of the global commercialized transgenic crops, select seven exogenous genes which basically cover the total commercialized crops, namely CaMV35S and FMV promoter, NOS terminater, mark gene NPTII, and aim genes PAT, EPSPS and CryIA(b).Use endogenous 18SrRNA gene as collate, design a large pairs of specific primers, screen the optimum primers groups, optimized the test condition and parameters, establishing the qualitative PCR detection system. On the basis of the mentioned above and the real-time PCR principal, we have designed and optimized the primers and probes, set up the real-time qualitative PCR detection system. Meanwhile, using the endogenous zein-maize and lectin-soybean as interior collate gene, EPSPS and CryIA(b) as target genes, set up the real-time quantitative PCR detection system.Transgenic integration site detection is the efficient method to identify whether the transgenic crops are approved to importor export. Some quantitative detection methods were reported in 2000 and 2001, and that is about Bt11 and Mon810 corn. Real-time 5' -nuclease PCR had been previously used successfully to determine quantitatively Roundup Ready(RR)soy and Btl76 maize in food. In this study, using BioSCien tomato as test material and the flanking region of the BioSCien insert sequence was determined using inverse PCR and real-time PCR..By means of verification, this sequence is specific for the transfection event. We set up the real-time PCR system for the quantitative event-specific detection of the genetically modified BioSCien tomato.Now the major results in the paper are abstracted as follows: 1.We collect 17 kinds of species which contain 75 varieties of global commercialized transgenic crops and the transgenic ingredients had been analyzed and confirmed basically through references and web site.2. The genomic DNA purification method is different slightly in different species of plants. We had optimized the efficient quick extraction method of CTAB, SDS and adopt suitable KIT and extracted the different plant genomic DNA successfully suited to PCR and real-time PCR system.3. By using plant endogenous 18SrRNA gene as target, specific primers and probes were designed and optimized. After that the condition and parameter were optimized. We established the genomic DNA quality identification system for PCR and real-time PCR.4. At the first time, we use CaMV35S and FMV promoter, NOS terminater, mark gene NPTII, and aim gene PAT,EPSPS and CryIA(b) genes as collate, design pairs of specific primers, screen the optimum primers groups, optimized the test condition and parameter, establishing the qualitative PCR detection system, the sensitivity is 0. l%(w/w).5.In this study,we have cloned CaMV35S and FMV promoter, NOS terminater, mark gene NPTII, and aim gene PAT, EPSPS and CryIA(b) genes used as positive collate.6. In this study, establishing contamination free PCR amplification system by adding UNG enzyme.7. At the first time, the reporter dye, FAM was linked to the 5' -end of the oligonucleotides of the probes, and the TAMRA was located at the 3' -end as quencher dye. We use CaMV35S and FMV promoter, NOS terminater, mark gene NPTII, and aim gene PAT, EPSPS and CrylA(b) genes as target sequences, design pairs of sp...
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