Construction Of The Testing System Of Multiplex Fluorescence PCR Method For Detecting Transgenic Organism

The most of transgenic organisms were transformed by vector, whichcontained 35S promoter derived from Cauliflower Mosaic Virus (CaMV) and Nos terminator derived from Agrobacterium tumefaciens nopaline. The most common cloning vector were pBR322 and its derivatives, which contained resistant gene of ampicillin or kanamycine with the aim for screening. The four markers of P-35S, T-35S, T-NOS and bla (or nptII) had become applied widely in screening genetically modified materials.To explore multiplex fluorescence PCR method for detecting transgenic organisms, P-35S gene and T-NOS gene were chosen as markers. According to the specific sequence of P-35S and T-NOS which had been used in transgenic crops frequently, two paires of primers and two paires of corresponding fluorophore double-chain probes were designed and synthesized for detection of two kinds of target genes simultaneously with MF-PCR in a tube. The standard plasmids and transgenic soybean samples were tested with the above method. The results showed that the described method was available.