Identification Of Flanking Sequences And Event-special Detection Of Transgenic Cry1Ab Rice

Transgenic plant has provided excellent resistance to many kinds of diseases,insects and advertise situations with new biotechnologies in recent years. Thetransgenic research also gives good chance to other problems. The physiological andbiochemical characters might be affected by the random insertion of target gene, aswell as the related metabolic pathway. The consequently unexpected effect might bethe possible threat to the safety of food. So, many countries published the rules for themanagement of gene-modified plants, and perfected the technology for the detectionof GM food, because more and more attention has been paid to the worry. In thepresent research, the location of target gene inserted was identified on the ricechromosome, and the expression of Cry1Ab was also analyzed in the44plantswithout selected gene. we hope that our research would provide useful information forthe further study of the metabolic pathway in transgenic rice and the necessaryevidence for the safety evaluation。We through improved thermal asymmetric interlaced PCR (TAIL-PCR), using fivesimple primers to amplified Cry1Ab gene on the44transgenic rice strains. The resultsshowed that we successfully separated and determinated the flanking sequences of36strains, the success rate about82%.Then we employed bioinformatics tools to analysisthe molecular characteristics of the exogenous gene of this transgenic rice. The resultsindicate that these strains can divide into four categories.24strains of them are belongto the first transformation event as their integration sites located in the OSC2;3strains are belong to the second transformation event as their flanking sequences arehighly homologous to the rice genome sequence of several different chromosomes;6samples, which harbored multiple copies of the transgene, are belong to the thirdtransformation event;3samples are belong to the fourth transformation event as theyhave tandem repeat of the transgene.These flanking sequences obtain from this study,which as essential experimental basis, are helpful to research and evaluate the potentialeffects that may be non-anticipatory.Transgenic Cry1Ab rice Bt01has applied forenvironmental release trials, and the southern hybridization experiments had shownthat it is single copy of the transgene.We establish the specific qualitative andquantitative detection method for the transformant of the transgenic Cry1Ab rice Bt01. The experiments showed that the minimum copy of transformants that can bedetective by this specific qualitative PCR system is ten, the LOD and the LOQ ofquantitative PCR detection system are5and10copies, respectively. In summary, theevent-specific detection method that establish by present study are applicable toqualitatively and quantitatively determine the exogenous Cry1Ab gene on thetransgenic rice Bt01. In addition, this method has credible stability, high specificityand sensitivity. It not only strengthens the GMO detection system, but also provides agood case for the detection of other genetically modified crops.