Research Of Producing HBV Vaccine With Transgenetic Tobacco

Hepatitis B is a world wide infectious disease which is induced by HBV (Hepatitis B virus). There are 280 million patients with HBV in the world. China is a high infectious zone with 20% people infected by HBV. HBV vaccination is the best way to prevent the infection of HBV, but the vaccine used now has many disadvantages. Now producing HBV vaccine with transgenic plant can overcome the traditional vaccine's shortcoming.In this experiment, we isolated HBV genome DNA from the serum that was obtained from the patient who infected HBV, and designed a pair of primers with Kozak consensus sequence based on the sequence AY040804 obtained in Genbank. From which, we amplified HBsAg gene that coding hepatitis B virus surface antigen S protein. By BLAST, we confirmed 1 base in this sequence was mutated. Perhaps it's a kind of new virus strain, and the sequence was submitted to Genbank, it is processing now.Moreover, we constructed a new plant expression vector-pBI121-HBsAg with HBsAg gene and pBI121 binary vector. pBI121-HBsAg contains CaMV35s promoter, nos terminator, Kan marker gene and HBsAg gene, and this plasmid was transformed to agrobacterium LBA4404 by freeze and thaw method, and then the tobacco leaf disc was transformed by LBA4404.Through this experiment, we want to establish a high efficiency transform system for tomato and tobacco; use CaMV35s high efficiency promoter to promote expression of HBsAg in tobacco, obtain regeneration plants contain specific protein with immunocompetence, and build a foundation for the further research of transgenic plant based vaccine.