Transformation Of Tomato With HBsAg/Truncated HCV Core Protein Fusion Gene

This research was to explore the feasibility of expressing HBsAg/HCV core protein fusion gene in plants for the purpose of developing a new type of plant-derived hepatitis bivalent edible vaccine.Hepatitis B Virus S gene was connected to the 5' end of Hepatitis C Virus core protein sequence with the technique of recombinant PCR,a designed DNA sequence which encoded a polypeptide linker (Gly₄Ser)₂ was used to splice the two fragments. DNA sequencing results showed that the fusion gene(BC) was successfully constructed.The fusion gene(BC),approximately1237bp,was cloned to the plant binary expression vector pBin438.The recombinant plasmid pBin438BC under the control of DE35S promoter was constructed and transformed into Agrobactrium tumefaciens EHA105 via freeze-thaw method. Inoculated explants from cotyledons and hypocotyls were cultured successively on preculture medium, co-cultive medium, selective medium and root-growth media to establish the optimize heredity transformation system, 9 Km-resistant tomato plants were obtained .The regenerated tomato plantlets were analyzed further by PCR and PCR-Southern blot.The results confirmed that the fusion gene had been integrated into tomato genome.