The Study On Detection Methods Of GMOs

Genetic modified crops have been applied in agriculture commercially in many countries due to theirs good characteristics such as high resistance to pest and weed. Many countries zealously develop the genetic modified technology, while at the same time they strengthen the monitoring and surveillance of genetic modified products with the increase in understanding of safety of genetic modified organisms. As China and many other countries adopt the labeled system towards the genetic modified crops, it's of great significance to develop the rapid detection techniques and their products- detection kits of GMOs (genetic modified organisms). This study aimed to study the key technologies and scientific issues related to establishment of rapid detection kits of GMOs.A pair of prime was designed to magnify the sequence of the S-enolpyuvyl-3-phosphoshikimic acid synthase enzyme (EPSPS synthase) of soybean by PCR according to the conservative sequence existing in the glyphosate tolerant plants. A probe linked to DIG was synthesized using the obtained DNA segment, and the effect of the ratio of target sequences over probes on hybridization efficiency was also studied, as to provide the scientific basis for establishment of the detection kit at DNA level.Through the conjugation between magnetism articles and Bt Cry lAc antibody, a RIA kit for detection of Bt Cry 1 Ac protein was set up, with shorter reaction time and simplified operation steps compared to normal detection methods using PCR.Two hybridoma cell lines, which secrete monoclonal antibody of Bt CrylAc protein stably after incubation many times, were obtained Their average chromosome number was 96. The monoclonal antibody, characterized as IgG1 isotype, reacted high specifically with Cryl Ab protein with sensitivity of 1.0ng/ml, and the across action rates, that caused 50% binding inhibition, with CrylAb and Cry2A, were 0.25% and 0.1%, respectively. One of lines was injected into rats to produce ascites, about 4-6mg/ml protein in the ascites was obtained, its affinity constant was 1.0928× 10101/mol. The antibodies secreted by these two cell lines have very high liters, which could be served as an important component of rapid detection strips.