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Virologica Sinica, 17 (1) : 60-65, 2002
Research Article
Study on Expression and Location of Hepatitis C Virus NS5A Protein in Insect Cells
Institute of Virology,Wuhan University.Hubei,Wuhan 430072,China
 Correspondence:
(234.26KB)  
Abstract
Full—length NS5A gene of Hepatitis C Virus was amplified by PCR.using plasmid pBAC25
containing HCV nonstructural protein genes as template.The amplified fragment(about 1.34kb)was
cloned into transfer plasmid pBuleBacHisA.The recombinant plasmid DNA was cotransfected with
genome DNA of wild type AcNPV to SF-9 il~ect cells.The recombinant baculovirus AcNS5A was ob—
rained by plaque selection.Restriction analysis and PCR identification indicated that the NS5A gene
was integrated into the genome of AcNSSA.and HCV NSSA gene was expre~sad as a 64kD protein in
SF·9 cells infected with AcNS5A.This protein can he recognized by anti.NS5A antib0dy when idend—
fied bv Western—blot.The Immune fluorescence assay and immune histo-chemistry assay were used tO
study the distribution of NS5A protein inside the cel1.Distribution of NS5A protein are changed with
the time in the recombinant virus infected SF一9 cells.Ns5A protein are changed with the time in the
recombinant virus infected SF-9 cells.NS5A protein iS 1ocated at the cel1 membrance at 24 hours post
infection and located at both the membrance and the nucleus after 48 hours.and ful1 of the cells after 72
hours.The result suggests that NSSA protein Was located on plasma membrance and in nucleus.it may
play an important role in RNA replication and in controlling the expression of the cell gene~during the
replication of the virus.
  Published online: 15 Feb 2002
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