利用昆蟲桿狀病毒系統表現B型肝炎核心缺損變異基因抑制B型肝炎病毒繁殖之功效(2/3)

A handy in vitro viral replication system is mandatory for hepatitis B virus (HBV)
study. A recombinant baculovirus with 1.3XHBV DNA construct was previously
designed to infect HepG2 cells. We adapted this system and set up another one
using 1.5XHBV DNA construct to generate our recombinant baculovirus, and we use
Huh7 cells instead of HepG2 cells to establish this system. HBV genome was
inserted into the baculovirus by recombination and the novel HBV recombinant
baculovirus was identified by enzyme digestion. The viral stock was purified and its
titre was determined. We then use the HBV recombinant baculovirus to infect Huh7
cell culture and demonstrate its ability to produce HBsAg. The produciton of
HBsAg was first detected in the media three days after infection. Its production was
in proportion to the loading amount of HBV recombinant baculovirus. A sustained
HBsAg production could be achieved by superinfection of this recombinant virus to
the already infected Huh7 cell culture. This system can be applied to the basic and
clinical studies of HBV.