[A preliminary study using RNA interference technique against replication of HBV in vitro].
Dachuan Cai, Yan Zeng, Youguo Li, Hong Ren
Abstract
Dachuan Cai, Yan Zeng, Youguo Li, Hong Ren
Abstract
OBJECTIVE: To identify the siRNA interference ability for the replication of HBV. METHODS: Based on the sequence of HBV in HepG2 2.2.15 cells in GenBank, one sequence targeting the C antigen of HBV was cloned into the RNA polymerase III based expression vector pSuper. This recombinant was electroporated into HepG2 2.2.15 cells and the expression of HBsAg and HBeAg was assayed using ELISA. RESULTS: The construction of the recombinant expression vector pSuper-C and its control vector pSuper was successfully confirmed by the results of enzyme digestion, electrophoresis and sequencing. However, there was no difference between the expression of HBsAg and HBeAg in the supernatant of HepG2 2.2.15 cell culture in the experimental and control groups. CONCLUSIONS: The constructed pSuper-C did not show an interfering effect on the replication of HBV in HepG2 2.2.15 cells. In order to display this effect, further study is needed
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OBJECTIVE: To identify the siRNA interference ability for the replication of HBV. METHODS: Based on the sequence of HBV in HepG2 2.2.15 cells in GenBank, one sequence targeting the C antigen of HBV was cloned into the RNA polymerase III based expression vector pSuper. This recombinant was electroporated into HepG2 2.2.15 cells and the expression of HBsAg and HBeAg was assayed using ELISA. RESULTS: The construction of the recombinant expression vector pSuper-C and its control vector pSuper was successfully confirmed by the results of enzyme digestion, electrophoresis and sequencing. However, there was no difference between the expression of HBsAg and HBeAg in the supernatant of HepG2 2.2.15 cell culture in the experimental and control groups. CONCLUSIONS: The constructed pSuper-C did not show an interfering effect on the replication of HBV in HepG2 2.2.15 cells. In order to display this effect, further study is needed
Key concepts: HBsAg, HBeAg, Molecular biology, Recombinant DNA, RNA interference, Virology, Biology, Vector (molecular biology)