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Virologica Sinica, 14 (2) : 135-139, 1999
Expression of E2 Glycoprotein Gene of Hepatitis G Virus in Insect Cells
Departmen~of Microbiology,Second Military Medical Uniwersity,Shanghai 200433
Hepatitis G virus E2 glycoprotein gene was amplified by polymerase chain reaction, and was inserted into baculovirus expression vector pF AST B AC HTa, constructing a recombinant transposing vector pF AST B AC E2. The plasmid pF AST B AC E2 was transformed into DH10B AC competent E. coli cells. High molecular weight DNA was prepared from the overnight cultures from the selected E. coli colonies, which was recombinant baculovirus shuttle vector containing HGV E2 gene, named Bacmid E2. The Bacmid E2 was transfected into Spodoptera fragiperda (Sf9) cells to get the recombinant virus. Fresh insect Sf9 cells and larvae of Spodoptera exigua were infected with the recombinant virus to express the target protein. The E2 glycoprotein recovering from the Sf9 cells and hemolymph cells of larvae of Spodoptera exigua exhibited a molecular mass of approximataly 54000D. Purified HGV E2 glycoprotein using affinity chromatography was used to develop an ELISA for detection of HGV E2
  Published online: 5 Jun 1999
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