Cloning of the non-structural gene 3 of hepatitis C virus and its inducible expression in cultured cells.
AIM:To study the inducible expression of hepatitis C virus ns3 gene (HCV ns3) in eukaryotic cells.METHODS: The ns3 gene was obtained from plasmid pBns3 by polymerase chain reaction and inserted into the cloning vector pGEM-T. Then, the ns3 was subcloned into the vector pMSG to generate dexamethasone (DM)-inducible expression plasmid pMSG-ns3.CHO cells were transfected by pMSG-ns3 using calcium phosphate precip-itation method and cultivated for 12h-24h. The transfected cells were induced with DM and the transient expression of NS3 protein was analyzed by ELISA and Western-blot methods.RESULTS:After treated with 3X10(-8)mol/L DM, the expression of NS3 was observed in the transfected CHO cells. A slightly higher level of NS3 was shown along with the time of DM treatment.CONCLUSION:The inducible expressing vector pMSG-ns3 might be helpful for further studies of the characteristics of the ns3 gene in vivo.
Zhang
Shu-Zhong
SZ
Liang
Jia-Jing
JJ
Qi
Zhong-Tian
ZT
Hu
Yi-Ping
YP