Studies On The Genetically Modified Papaya And Tomato Using Molecular Technology Methods

Since the first genetically modified crop tobacco came into being in the world in 1983, the genetically modified technology has had a profound influence on the yield and quality of the crops and created enormous economic and social benefits. However, genetic engineering may lead to unpredictable plant property changes that even difficult to be aware in short-term, may bring the enormous risks for environment, ecosystem, personal health and so on. In order to strengthen the control of transgenic crops and products, many countries have issued their own managements and policies. Thus, the detection for genetically modified crops is urgently needed. Based on that, this research has established the methods of common PCR and Real-time fluorescence PCR respectively of genetically modified papaya and tomato, The main contents of the paper are as follows: Genetically modified papaya and tomato common qualitative PCR detection technology1. Selected ribulose bisphosphate carboxylase/oxygenase large subunit gene (RBCL gene) as endogenesis reference gene and design universal primes, choosed papayas, tomatoes and strawberrys as the experiment materials . The result indicated: RBCL gene can be regarded as endogenous gene in the genetically modified papaya and tomato.2. Aimed at the common extraneous sequence of genetically modified papaya and tomato: CaMV35S promoter sequence, the NOS termination sequence, the NPTII gene, the GUS gene, the CH5B gene, the OSMOTIN gene, the PRSV-CP gene and the PRSV-RP gene, then designed separately specially primes, acted as the ordinary qualitative PCR examination research, and optimized the PCR system.3. Designed primes of 35S/NOS and 35S/CP, carried out a pair primes to check 2 or more of the extraneous fragment, and optimized the PCR system.4.Established dual and trial qualitative PCR detection methods, and optimized the PCR system.5.Cloned and sequenced the PRSV-CP gene, the PRSV-RP gene, the CH5B gene, the OSMOTIN gene, the GUS gene, extracted their plasmids as the positive template; and also confirmed different boundary connection sequence in 35S and the PRSV-CP gene between genetically modified papaya of the PRSV-CP strains.