Document Type: Research Paper
Institute of Biochemistry and Biophysics, Tehran University, Tehran, Iran.
Hepatitis B virus (HBV) is a serious global health problem. The development of a safe and effective vaccine
would help infection prevention. Previous hepatitis B vaccine production involved the isolation of the noninfectious particle from chronic HBV carriers. DNA recombinant technology has been used for vaccine
production without having been contaminated with blood-born infectious agents. Vaccine production in
mammalian cells has the advantage of being correctly modified and folded in comparison to other lower
hosts. The surface protein coding genes, S (Major protein) and pre s2+s (Middle protein) of hepatitis B virus
(HBV), were amplified from the mother plasmid containing the adr serotype virus genome. The s and pre
s2+s amplicons were separately cloned in pBlueskript IIks(+) vector as pNM-sa2 and pNM-Psa2 intermediates
respectively, then released and recloned in pcDNA3 mammalian expression vector. The correct pNM-Sb2
and pNM-Psb2 constructs containing s and pre-s2, respectively, were used to transfect the mammalian
Cos-7 cell line. The major and middle proteins were secreted by this cell line and collected from the culture
medium. Some features of gene cloning strategy and expression of these proteins are discussed.