Abstract:In order to efficiently identify suspect transgenic positive samples in honey, ten transgenic promoters and terminators, i.e., CaMV35S promoter, NOS terminator, FMV35S promoter, NOS promoter, Ubi promoter, TA29 promoter, E9 terminator, OCS terminator, g7 terminator, CaMV35S terminator, were used as research targets. A series of experiments were conducted for these ten promoters and terminators, including primers combination screening, reaction condition optimization, sensitivity determination, and test comparison of quality control products. Three groups of multiple real-time fluorescence PCR reaction systems were established and applied to the detection of commercial honey. The result showed that the established three groups of multiple real-time fluorescence PCR reaction system could accurately detect the target gene, with sensitivities of 0.01, 0.04, 0.04 ng/μL, respectively. The tests of 14 genetically modified quality control products showed that the detection results of multiplex real-time fluorescence PCR and single-plex real-time fluorescence PCR were consistent. The screening of 40 commercial honeys showed that two honeys contained pCaMV35S and tNOS, and the others were not detected, greatly improving the detection efficiency. This method could provide a fast screening and detection technology for the detection of genetically modified ingredients in honey, and meanwhile, it could also be applicable to the detection of genetically modified ingredients in other products.
