Hepatitis B Virus HBcAg Gene Transformation in Escherichia coli DH5a Using pEGFP-N1 Dini Cahyani 1) , Lalu Unsunnidhal 2,3) , Tri Untari 4) and Asmarani Kusumawati 5,6)
1 Master Program of Biotechnology, Graduate School of Universitas Gadjah Mada, Jalan Teknika Utara, Kocoran, Caturtunggal, Depok, Sleman, Yogyakarta 55281 Indonesia
2 Food Technology Study Program, Faculty of Food Technology and Agroindustry, University of Mataram, Mataram, 83125, Indonesia
3 Biomedical Field, Nursing Study Program, STIKES Yarsi Mataram, West Nusa Tenggara 83361, Indonesia
4 Departement of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
5Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
6Biotechnology Study Program, Graduate School of Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
Abstract
Hepatitis B virus is a virus that attacks humans and it can cause liver cancer. Hepatitis B subgenotype B3 is commonly found in Southeast Asia. Thus, the development of a vaccine is carried out to prevent the spread of the Hepatitis B virus in Indonesia. The purpose of this study was to transform the hepatitis B virus HBcAg gene using the pEGFP-N1 vector on Escherichia coli DH5a as host cells. The method in this research was codon optimization and plasmid transformation of recombinant DNA in pEGFP-N1. The recombinant cloning was divided into two stages, that was making competent cells and transforming recombinant DNA plasmids. The HBcAg gene was obtained from the results of isolation and cloning that had been carried out by previous researchers, would be constructed on the pEGFP-N1 vector. The results obtained in this study were that the HBcAg gene was successfully cloned and transformed into Escherichia coli DH5a (pEGFP-N1-HBcAg) by heat shock method using MgCl2-CaCl2 reagent. Based on the results of PCR colonies which stated that the transformant bacteria E. coli DH5a succeeded in carrying the pEGFP-N1-HBcAg plasmid marked by the presence of DNA bands with a length of 152 bp. Therefore, the construction of pEGFP-N1-HBcAg using Escherichia coli host cells can be used as a DNA vaccine candidate.
Keywords: DNA vaccine, Escherichia coli DH5a, Hepatitis B, pEGFP-N1-HBcAg
