Establishment Of The Detection Method For Identification Of GMOs Utilizing PCR

With the rapid development of genetic engineering technology in recent years,transgenic technology has become successful.The GMOs and their products are increasingly involved in the food and biotechnology industry and issues surrounding their safety have attracted more and more attention.Policies and regulations relevant to GMOs have been put into practice to enhance management of GMOs and their products.By the end of 2002,There have been more than 40 countries and regions taking labelling management voluntarily or mandatorily to supervise GMOs in the world.The premise to implement these laws and regulations is to establish a stable and accurate technology to identify GMOs.Currently,the detection of GMOs and their products mainly based on PCR technology,and its derived quantitative competitive PCR,real-time quantitative PCR etc..PCR have so many advantages like specificity,high sensitivity,rapidness,accurateness and repeatability that almost countries take this method in GMOs detecting programs.In this study,transgenic reference material were taken as templates in qualitative PCR,quantitative competitive PCR,and fluorescence quantitative PCR respectively to test precision of PCR.The findings are as follows:1.Collection of the background information of known commercialized genetically modified crops worldwide by visiting websites,databases.Nineteen accessions of GMOs were collected successfully, of which eight were purchased from the transgenic reference library in the Europe(Fluka,Buchs, Switzerland).2.To improve the efficiency in detecting GMOs,a high-throughput PCR detection method was established by optimizing PCR parameters in detection of exogenous genes integrated in transgenic soybeans references.3.To improve the sensitivity of PCR,event Bt11,TC1507,Bt176,MON863,MON810,GA21, CBH-351 and the corresponding exogenous genes in introduced transgenic maize references were detected,of which 6 transgenic events reached a low limit of 0.1%,the highest level in existing detection criterions.4.Based on the GMOs detection technology system established,stable PCR detection programs were set up for identification of transgenic insect-resistant rice and cotton,as well as transgenic sugarcane to resist mosaic disease and drought,which provide references for establishment of relevant detection standards.5.Construction of the non-homologous template CaMV35S competition controls,the establishment of transgenic soybeans CaMV35S promoter quantitative detection system,and the qualitative PCR detection of GMO products,the low detection limit of 1%to basically meet the current minimum of transgenic logo needs.6.A fluorescent dye SYBRGreen I to replace expensive Taqman probe,the establishment of a transgenic herbicide-resistant soybean and TC1507 transgenic insect-resistant maize fluorescence quantitative PCR detection system,the use of transgenic reference established by the linear regression equation,through known reference.As the known sample to verify the results entirely correct,and the detection sensitivity of the two systems reached 0.04 percent,the international community has fully satisfied the quantitative detection of transgenic needs.