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. 2003 Jun;77(12):7017–7025. doi: 10.1128/JVI.77.12.7017-7025.2003

FIG. 3.

FIG. 3.

Expression of retroviral packaging components by recombinant vaccinia viruses shown by Western blotting. CV-1 cells were infected with the indicated viruses, and total cellular extracts were prepared and subjected to the Western blot procedure. (A) Incubation of the blots with a gag-pol antiserum revealed a characteristic pattern of protein bands, including the Pr65 gag-specific band and the p30 capsid (CA) band (arrowhead, right side) in the packaging virus and two clones of the triple viruses. Lane 1, protein markers. Lane 2, positive control, lysate of cells infected with the virus vHA-MLVg, which has a single insert of the gag-pol sequences (see text). Lane 3, negative control, lysate of cells infected with WR wild-type virus. Lane 4, lysate of cells infected with the packaging virus. Lane 5, lysate of cells infected with triple virus clone 1. Lane 6, lysate of cells infected with triple virus clone 2. (B) Incubation of blots with VSV-G antibodies revealed the typical 55-kDa band of the VSV-G glycoprotein in the packaging and triple viruses. Lane 1, protein markers. Lane 2, positive control, lysate of cells infected with the virus vDD4-mH5-VSVg, a recombinant WR-based virus with a single VSV-G gene insert. Lane 3, negative control, lysate of cells infected with wild-type WR virus. Lane 4, lysate of cells infected with the packaging virus. Lane 5, lysate of cells infected with triple virus clone 1. Lane 6, lysate of cells infected with triple virus clone 2. Numbers at the left indicate the size of the protein markers, and those at the right show the size of characteristic bands (in kilodaltons).