Reserach On Detection Of Transgenic Rice With Bar, Bt Gene And Degradation Of Transgene DNA

With the deterioration of the environment and the shortage of natural resources, there is a food crisis spreading around the world which makes the transgenic technology develop rapid and get application in the Agricultural production. Nowadays, more and more transgenic productions already got into the commercialization stage as well as the people’s food chain directly or indirectly. Governments have published a series of laws and regulations which aim at transgenic organisms because of the widely public concerns about the effection of the transgenic crops and its products to the people health and ecological environment.Detection is the premise of the supervision, this experiment explore the detection methods of transgenic rice with Bar and Bt gene. Degradation of endogenous and exogenous gene of transgenic rice in the course of rice product processing was also researched in this paper. The details are as follows:Based on the DNA sequences of rice endogenous gene SPS and exogenous gene (Bar, Bt, CaMV35S promoter, Ubiquitin promoter, NOS terminator), specific primers were selected to establish two triplex PCR (Polymerase Chain Reaction) detection systems for transgenic rice with Bar and Bt gene, which could provide technology support to the detection of transgenic rice、safety estimation along with the Traceability research. Results showed that the first triplex PCR system can simultaneously detect SPS, Ubquitin and Bt gene; the second can simultaneously detect CaMV35S, NOS and Bar gene.The reaction condition of two triplex PCR detection systems is as follows:the sample solution was kept at94℃for5min, subjected to40cycles of amplification (94℃for1min,57℃for1min, and72℃for lmin), and kept at72℃for7min. According to the detection results, the sensitivity value of each system was0.5%. In the meanwhile, the addition volum of each primer and the reaction condition of two systems were same, which simplify the operation.In this study, sweet rice wine and rice cracker were made by using transgenic rice as main material. The degradation of SPS gene, CaMV35S promoter, Ubiquitin promoter, NOS terminator, Bar gene and Bt gene were detected by using different fragments-size PCR primers. It was found that the stability of genes was different under different processing conditions. After four days’fermentation, the largest detectable fragments of SPS, CaMV35S, Ubiquitin and NOS, Bar and Bt gene in the sweet ferment rice were916,446,133,180,370and635bp, respectively. The stability of the genes in ferment was found to be in the following decreasing order:NOS terminator, Bar gene, CaMV35S promoter, Bt gene, SPS gene and Ubiquitin promoter. The Severe degree of the five different hot processings was found to be in the following decreasing order:frying, baking, microwave treatment and common boiling. The result of high-pressure boiling is the same with common boiling. The stability of the genes in five different hot processings was found to be in the following decreasing order:CaMV35S promoter, NOS terminator, Bar gene, Ubiquitin promoter, Bt gene and SPS gene. The fragments whose length is under200bp are characteristic of good stability.