Cloning and Expression of HBcAg Using Food Grade Vector pNZ8149 in Lactococcus Lactis for the Development of HBV Therapeutic Vaccine
- DOI
- 10.2991/978-94-6463-112-8_76How to use a DOI?
- Keywords
- HBV; CHB; HBcAg; pNZ8149; usp45; Lactococcus lactis NZ3900; nisin
- Abstract
Hepatitis B virus (HBV) infection remains a major cause of chronic liver diseases which may develop into cirrhosis and hepatocellular carcinoma (HCC). Although HBV prophylactic vaccine is available, currently there is no complete cure for chronic hepatitis B (CHB). One of the alternatives for CHB treatment is therapeutic vaccine. The HBV core antigen (HBcAg) becomes a suitable candidate for HBV therapeutic vaccine since it is known as a potent inducer of B-cell and T-cell responses. In this study, we report the development of HBV therapeutic vaccine by expressing HBcAg derived from HBV subgenotype B3, which is one of the predominant sub-genotypes in Indonesia. A food grade expression vector pNZ8149 which contains nisin-controlled gene as an inducible promoter, was used to express HBcAg in Lactococcus lactis NZ3900. Moreover, signal peptide usp45 was added in the N-terminal of the gene of interest to increase protein secretion efficiency. The native HBV core gene was synthesized and successfully cloned into plasmid pNZ8149 and transformed into Lactococcus lactis NZ3900 using lactose as a selection marker. The expression of HBcAg was induced using 10 ng/mL nisin and confirmed using dot blot hybridization analysis. Finally, the target protein which is 21 kDa in size was obtained by partial purification using size exclusion chromatography. In conclusion, HBcAg was successfully cloned and expressed using a food grade expression vector pNZ8149 in Lactococcus lactis NZ3900. These findings will pave the development of therapeutic vaccine for CHB treatment in the future.
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- © 2023 The Author(s)
- Open Access
- Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.
Cite this article
TY - CONF AU - Rifqiyah N. Umami AU - Rahma I. Anwar AU - Hidayah Murtiyaningsih AU - Apon Z. Mustopa PY - 2023 DA - 2023/03/01 TI - Cloning and Expression of HBcAg Using Food Grade Vector pNZ8149 in Lactococcus Lactis for the Development of HBV Therapeutic Vaccine BT - Proceedings of the 1st International Conference for Health Research – BRIN (ICHR 2022) PB - Atlantis Press SP - 833 EP - 846 SN - 2468-5739 UR - https://doi.org/10.2991/978-94-6463-112-8_76 DO - 10.2991/978-94-6463-112-8_76 ID - Umami2023 ER -