CLONING OF BFV3026 GAG GENE AND EXPRESSION IN E.coli

With polymerase chain reaction (PCR), a fragment encoding the Gag protein was amplified from the Hirt DNA of bovine foamy virus3026-infected cells, which was isolated previously from the peripheral blood mononuclear cells of an infected cattle in Tianjin.. Sequence analysis revealed that it had high identity with that of the reported BFV in GenBank. Then this gag gene was ligated into plasmid of pET 32A for prokaryotic expression. BL21 (DE3) of E.coli transformed with the recombinant plasmid of pET gag was induced to specifically express the Gag protein in high level. The expressed product was characterized by SDS-PAGE and Western blot analysis, which occupies 40% of total bacterial protein. The report has laid a basis for searching the possible function of Gag in virus assembly.