CHITOSAN AS DELIVERY SYSTEM FOR HBcAg GENE EXPRESSION WITH pEGFP-C1 ON HELA CELLS

Hepatitis B is a viral liver infection caused by hepatitis B virus (HBV), which may lead to chronic liver disease, cirrhosis, and liver cancer. Recombinant DNA technology offers a promising approach for developing innovative hepatitis B vaccines. In this approach, the gene encoding hepatitis B core antigen (HBcAg) can be introduced into Escherichia coli to produce recombinant antigenic proteins. These proteins then formulated with nanoparticles as vaccine delivery systems. The aim of this study was to determine the effectiveness of chitosan nanoparticles as a delivery system for HBcAg VHB gene expression using pEGFP-C1 in HeLa cells as eukaryotic host cells. The methods used in this study included reculturing transformant bacteria obtained from previous research, isolating recombinant DNA plasmids, performing plasmid restriction analysis and PCR, sequencing, transfecting HeLa cell cultures, and observing EGFP-HBcAg protein expression through fluorescence microscopy. PCR analysis confirmed that the transformant E. coli DH5α successfully harbored the pEGFP-C1-HBcAg plasmid. Restriction analysis of the recombinant plasmid produced two distinct DNA bands, indicating successful plasmid digestion. Following transfection of HeLa cells, green fluorescence was observed in cells treated with chitosan nanoparticles, suggesting expression of the EGFP-HBcAg protein. These findings indicate that chitosan nanoparticles have potential as a delivery system for hepatitis B DNA vaccines using the pEGFP-C1 vector