Study On High-flux Detection Methods To The Exogenous Genes In Genetically Modified Plants

Fallowing the abundance of the genetically modified product (GMP), people pay more attention to the safety of GMP. Currently, we haven't rendered reliable answer to the safety of GMP. It's necessary to develop and improve the detection methods for the safety control of GMP.At present, there are kinds of distinguishing detection methods. It's significant to develop a kind of functional, handy, effective detection method for the advancement of transgenic biotechnology and the safety control of GMP.According to the feature of exogenous genes in transgenic plants and combining the microtube hybridization, we established a group of functional, handy, effective detection method.Most of exogenous genes in genetically modified plants apply CaMV35S and NOS as their promoter and terminator. In this study, firstly we design pairs of primers according to the CaMV35S and NOS and amplify the plasmid pBI121 that contains CaMV35S and NOS by PCR and label the PCR products as probes. After clipping the nylon membrane into strips, the DNA of multi-samples is fixed on the strips, hybridized with probes and the hybridization signal is detected in 2ml microtubes. The result is that this method can detect multi-samples by one or two probes. At last it shows that the result of hybridization and PCR are unanimous after the hybridization signal is verified by PCR, and it shows that to detect exogenous genes in multi-samples is available by microtube hybridization.For a single transgenic plant, it contains not only the promoter and terminator, but also the goal-gene and mark-gene. In this study we use genetically modified cotton as the sample, and design a group of oligonucleotide probes according to the exogenous genes in genetically modified tobacco. Then the amination probes are fixed on nylon strips processed by EDC. The DNA of sample is hybridized with the oligonucleotide probes after being marked and the hybridization signal is detected in 2ml microtube. The result is that this method can detect multi-exogenous gene in one sample. At last the sample is detected by PCR with the primers of exogenous genes in genetically modified tobacco, and it finds that the result of hybridization and PCR are unanimous, and it shows that to detect multi-exogenous genes in one sample is available by microtube hybridization.Neomycin phosphotransferaseⅡ(NPTⅡ)gene usually used as labeled gene can be translated into RNA and be transcribed into protein. In this study we detect the RNA translated by the exogenous gene in multi-samples with NPTⅡprobe. Firstly we amplify the pBI121 that contains NPTⅡwith PCR and labeled the product of PCR to be probe. RNA samples are fixed onto the nylon strips, and then hybridized with the probe and the hybridization signal is detected in the 2ml microtube. It found that one probe can detect RNA translated by exogenous gene in multi-samples. To verify the hybridization signal, RNA samples of multi-samples are detected by RT-PCR with NPTⅡprimers. Then it finds that the result of hybridization and PCR are unanimous after the hybridization signal is verified by RT-PCR, and it shows that to detect RNA translated by exogenous genes in multi-samples is available by microtube hybridization.