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The Expression of Codon Optimised Hepatitis B Core Antigen (HBcAg) of Subgenotype B3 Open Reading Frame in Lactococcus lactis

  • Mustopa, Apon Zaenal (Research Centre for Biotechnology, Indonesian Institute of Science (LIPI)) ;
  • Wijaya, Sri Kartika (Research Centre for Biotechnology, Indonesian Institute of Science (LIPI)) ;
  • Ningrum, Ratih Asmana (Research Centre for Biotechnology, Indonesian Institute of Science (LIPI)) ;
  • Agustiyanti, Dian Fitria (Research Centre for Biotechnology, Indonesian Institute of Science (LIPI)) ;
  • Triratna, Lita (Research Centre for Biotechnology, Indonesian Institute of Science (LIPI)) ;
  • Alfisyahrin, Wida Nurul (Research Centre for Biotechnology, Indonesian Institute of Science (LIPI))
  • Received : 2018.06.22
  • Accepted : 2019.02.12
  • Published : 2019.09.28

Abstract

Hepatitis B treatments using immune therapy are gaining interest because of the improvements in dendritic cell performance for antigen presentation, which induces an appropriate immune response and raises patient survival rates. This research aims to produce a significant amount of the HBcAg antigen, which can induce an immune response and have a curative effect on HBV infection. In this study, the HBV subgenotype B3 of the HBcAg gene was used, which is dominant in Indonesia. Further, Lactococcus lactis bacteria was used as the host because of its safety and tightly regulated protein expression. The codon usage for the HBcAg gene was optimized to improve protein expression in L. lactis, which is important because a codon is not random between species. The HBcAg gene is attached to a pNZ8148 plasmid and transformed into the L. lactis NZ3900 expression host. The results confirm that a positive protein band (21 kDa) in two fractions of purified HBcAg was recognized by both western blotting and dot blot hybridization, even if the HBcAg optimized codon has higher GC contents than that suggested for L. lactis expression. Overall, this research strengthens the broad use of L. lactis bacteria for any protein expression, including higher protein expression of codon optimized HBcAg gene compared to non-optimized genes. Furthermore, the improvement in the codon optimization of the HBcAg gene significantly increases the total protein expression by 10-20%, and the expression level of the codon optimized HBcAg increases 1.5 to 3.2-times that of the native HBcAg.

Keywords

Lactococcus lactis;HBcAg gene;codon optimisation;protein purification;western blotting;dot blot hybridization

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