Anti-type ¢ñ Diabetes Gene Into Tobacco, Tomato

Type I diabetes mellitus is a T cell-mediated autoimmune disease target at pancreatic isletβcells, which is the comprehensive interaction of both genetic and environmental factors. Heat shock protein 65 (HSP65) and the fusion fragment HSP65-6×P277 both have distinctive effect toward Type I diabetes.In this study, we transferred sequence HSP65 and HSP65-6×P277 into tobacco leave and tomato cotyledons mediated by Agrobacterium EHA105 with Kanamycin as plant selection marker. The objective is to obtain transgenetic plants containing anti-diabetic plant vaccine which is biologically effective. Some preliminary progresses are as follow:1. Construction of plant expressing vector pCAMBIA2301-HSP65 and pCAMBIA2301-HSP65-6×P277 containing CaMV35s promoter with Kanamycin resistance selection marker.2. By electrical stimulation, we transformed the plasmid vectors pCAMBIA2301-HSP65,pCAMBIA2301-HSP65-6×P277 into Agrobacterium EHA105 successfully.3. The Agrobacterium strains containing pCAMBIA2301-HSP65-6 X P277 infect tobacco leave,43 kan-resistant plants were screened, in which,34 were positive.4. The total RNA was extracted from the leaves of positive tobacco seedlings RT-PCR showed the successful integration and expression of the transgene into tobacco genome.5. The Agrobacterium strains containing pCAMBIA2301-HSP65 and pCAMBIA2301-HSP65-6×P277 infected tomato cotyledons,11 and 8 positive plants were derived from kan-resistant selection.6. RT-PCR of the total tomato RNA results showed that 5 plants expressed 2301-HSP65 and 4 plants expressed vector 2301-HSP65-6P277 at RNA-level7. Genetic analysis and protein expression are undergoing.We obtained in this study transgenic positive tobacco and tomato plants with the transgenes, it's a basis to further understand the anti-reactor type 1 diabetes edible vaccine,and to establish a new way to controll the disease.