CLONING AND TRANSFORMATION OF HEPATITIS B SURFACE ANTIGEN(HBsAg) GENE TO TOMATO (Lycopersiconesculentum Mill.)

Hepatitis B virus (HBV) is one of the most serious infectious diseases among human races worldwide. It is estimated that nearly 5% of world population are chronic carriers of HBV. Derivation of the hepatitis B vaccines from yeast and mammalian cells are tedious and expensive, so it can limit the use of these vaccines in developing countries. Biological studies have revealed information that plants are known to act as bioreactors that might successfully be able to offer economical ways for the production and boosting of recombinant proteins. Production of vaccines via plant expression system seems to be effective and promising process which can help to mitigate laborious and cost effects. In this study the appropriate primers were designed concerning highly plant expression sequence at the both sides of HBsAg gene and adequate restriction sites. HBsAg gene was cloned in a plant expression vector; pCAMBIA1304, to deploy constructs which they then were introduced into Agrobacterium tumefaciens strain LBA4404 using freeze-thaw procedure and leaf disk technique for transforming into cotyledonary explants of tomato. Transgenic lines were screened and regenerated from selection media containing cephotaxim and hygromycin and CTAB method used to isolated genomic DNA of the transgenic plant. The use of PCR technique and sequencing at the further examination could detect and made a proof of the presence of HBsAg gene in the transgenic plant.