|Title:||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(3/3)||Authors:||倪衍玄||Keywords:||hepatitis B virus;baculovirus||Issue Date:||2004||Publisher:||臺北市：國立臺灣大學醫學院小兒科||Abstract:||
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 production 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.
We then introduced the recombinant baculovirus into the portal vein of the mice to test
the in vivo model. The histologic examination of the mice showed the
necroinflammatory changes. However, the typical groundglass pattern of HBV
infection found in human pathology was not demonstrated in the mice model. A
microarray study was done to compare the RNA expression in the mouse liver infected
with HBV-recombinant baculovirus and with baculovirus vector only for the future
|Appears in Collections:||醫學系|
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