RAMANJINI GOWDA, P. H.ASHA, V. NAIR2017-06-192017-06-192009-07-15Th-9333http://krishikosh.egranth.ac.in/handle/1/5810021798Hepatitis B virus (HBV) infection remains as a worldwide health problem. Immunization is the only means to prevent the occurrence of this disease. As the available vaccines are much expensive, there is still need for a less expensive vaccine source especially in most of the developing countries. Expression of vaccine antigens in plants provides a cheaper source of vaccine production. Also plant based vaccines can be used as oral vaccines. The present study lays emphasis on study of integration and stability of the recombinant protein expressed in tobacco plants. Restriction digestion analysis of the gene construct pHB118 with restriction enzymes EcoRI and BamHI yielded two separate bands of 9.7kb and 3.6kb size. The presence of HBsAg gene in putative transformants of the T2 generation of tobacco plants were confirmed by PCR analysis. All the tobacco putative transformants used for PCR analysis showed the presence of 900 bp band. Transgenic and control tobacco plants evaluated for the presence and expression of the hepatitis B surface antigen gene were confirmed by SDSPAGE, protein dot blot, ELISA and Western blot. The T2 generation seeds obtained from the transgenic tobacco plants were tested for the germination in presence of kanamycin. It was observed that the segregation ratio was 3:1 indicating mendelian inheritance. The observations on growth and yield parameters of T2 generation tobacco plants transformed with hepatitis B surface antigen gene were recorded. It was found that the presence of foreign gene did not inhibit the growth of the transgenic plants.ennullThesis