G. Renuga* and A. Babu Thandapani
Department of Biotechnology, Ultra College of Pharmacy, Madurai, Tamil Nadu, India
Edible vaccine might be solution to get rid of various ailments as it has more advantages compared to traditional vaccine. Vaccines are used most widely in the world also reduced mortality rate caused by various infectious organisms and their use has been considered one of the safe and effective measure and to control various infectious diseases. In the present study, high-level expression of recombinant Hepatitis B surface Antigen (rHBsAg) gene PreS2-S was achieved in yeast Pichia pastoris. A single copy of HBV M gene (PreS2-S) was cloned in PICZA vector thereby rHBsAg protein could then be expressed by induction with methanol. Transformation can be carried out using Agrobacterium T-DNA vector, high levels of expression PreS2S-21 (843 bp) can be achieved by using strong specific plant promoters. M-gene was amplified by PCR. The plasmids were transformed into potato callus via Agrobacterium tumefaciens. Potato calluses were developed as mass culture on MS medium supplemented with growth hormones. The production of vaccines in callus cells was analyzed by preparation of protein and immunized in to mice and confirmed the level of protein expression from potato callus by ELlSA. Purified rHBsAg protein size was confirmed by SDS PAGE and the molecules were visualized by using Atomic Force Microscopy and determined to be organized in 20-25 nm particles. Immunogenicity of purified rHBsAg proteins was demonstrated in mice which were shown to produce specific anti-HBsAb response. The use of plant derived vaccines may overcome some of the major problems encountered with traditional vaccination against infectious disease.
Keywords: Edible vaccine, Hepatitis B surface antigen, Pichia pastoris, PCR, Transgenic callus