Development of Transgenic Maize using Immature Embryos of HiII Genotype as a Vaccine Candidate

Abstract

Problem statement: Plant-based vaccines posses some advantages over other types of vaccine biotechnology such as safety, low cost of mass vaccination programs and wider use of vaccines for veterinary medicine. These study was undertaken to develop the transgenic maize as edible vaccine candidate for animals. Approach: The immature embryos of HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vector V622. The vector was harbored nptII gene, which confers resistance to paromomycin and ApxIIA gene was produced ApxII toxin, which was generated in various serum types of A. pleuropneumoniae as a target gene. Results: The 1,027 immature embryos were immersed for 5 min in the Agrobacterium solution and then these were co-cultured on solid co-cultivation medium at 28°C for 2 days. After the delay period, the scutellum explants, axis removed embryos, were cultured on medium with 50 mg L^-1 paromomycin for first 2 weeks and a paromomycin-resistant callus were sorted out on the selection medium with 100 mg L^-1 paromomycin for 4×14 days. A total of twenty callus clones were selected and sixteen-putative transgenic plants were regenerated. Among them, only five plants contained the integrated nptII gene, which was confirmed by Southern blot analysis. Conclusion: These results demonstrated that the nptII and ApxIIA genes integrated into the maize genome and that transgenic maizes can be use as vaccine candidate.