Expression of Hepatitis B Surface Antigen in Coleus forskohlii
By: Neha Guleria, Paramanahally Hanumanthe Gowda Ramanjini Gowda, Satish Kumar Kariyaiah, Renu Pasricha
Key Words: Agrobacterium, Coleus forskohlii, HBs Ag, VLPs, Western blot.
Int. J. Biomol. & Biomed. 5(1), 29-39, July 2016.
Hepatitis B, a common viral infection, affecting millions of people every year, leads to disability and death. It has become one of the alarming diseases in the world. Vaccination is the best possible way to prevent this deadly viral infection and its consequences. Unaffordability of the mass vaccination program due to low health budgets especially in developing countries have demanded the economical, effective and easily available vaccine production against hepatitis B virus. The expression of subunit vaccines and recombinant proteins in plants is a convenient, safe and potentially economical platform technology. Hence, development of a plant-based vaccine could be promising. Therefore, the present investigation focused on expression and large- scale production of hepatitis B surface antigen (HBsAg) in Coleus forskohlii for the development of vaccine. Eight transformed C. forskohlii plants were generated via Agrobacterium -mediated transformation method. The integration of 681bp of HBsAg gene into the plant genome was confirmed using PCR. SDS-PAGE showed the presence of ~48kDa dimer and ~24kDa monomer form of HBsAg protein and the expression of recombinant protein was further confirmed by western blot. C. forskohlii expressed HBs Ag was recorded 260µg/g leaf fresh weight as measured by ELISA. Transformed plants of HBsAg showed the accumulation of Virus Like Particles of 22 nm size using transmission electron microscopy. This study offers a great potential for the large -scale production of hepatitis B vaccine in C. forskohlii which provides a strategy for contributing a means to achieve global immunization for the hepatitis B prevention and eradication.