Open AccessCloning of the non-structural gene 3 of hepatitis C virus and its inducible expression in cultured cells
Author(s) -
Shu-Zhong Zhang,
Jia-Jing Liang,
Zhongtian Qi,
Yiping Hu
Publication year - 1999
Publication title -
world journal of gastroenterology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.427
H-Index - 155
eISSN - 2219-2840
pISSN - 1007-9327
DOI - 10.3748/wjg.v5.i2.125
Subject(s) - transfection , ns3 , microbiology and biotechnology , cloning (programming) , plasmid , expression vector , hepatitis c virus , western blot , chinese hamster ovary cell , biology , virology , vector (molecular biology) , gene , virus , cell culture , recombinant dna , biochemistry , genetics , computer science , programming language
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.