Genetic construction of HBsAg gene subgenotype B3 in Lactococcus lactis as hepatitis B vaccine candidate

Abstract

Hepatitis B is an inflammatory liver disease caused by HBV (Hepatitis B Virus). Hepatitis B surface antigen (HBsAg) induces immune system forming antibodies. HBV subgenotype B3 is common in Asian Countries. Thus, the development of HBsAg subgenotype B3 vaccine was done because its prevalence is high in Indonesia (especially in Javanese) and other Asian countries. The research methods were preparation of the HBsAg gene subgenotype B3, cloning and transformation the HBsAg gene in Escherichia coli MC1061, and transformation in Lactococcus lactis (L. lactis). HBsAg gene subgenotype B3 was obtained from the pIDT-HBsAg subgenotype B3 plasmid. The HBsAg gene subgenotype B3 successfully cloned and transformed into E. coli MC1061 and L. lactis. The PCR results of the transformant E. coli MC1061 (pNZ8148-HBsAg subgenotype B3) colonies were found in colonies 8, 17, and 20 indicated by the presence of 1226 bps bands. 8 colonies were obtained from PCR results of L. lactis transformants (pNZ8148-HBsAg subgenotype B3). The construction of the HBsAg subgenotype B3 gene has 100% similarity compare to the hepatitis B virus isolated from Java on 1839. Therefore, the construction of pNZ8148-HBsAg subgenotype B3 using host cells L. lactis can be used as a vaccine candidate.