FIG 2.
Genetic organization of PB1-, NP-, and M1-expressing pJSA1175 recombinant plasmids containing homologous sequences from vaccinia virus and the resulting protein expression in HeLa cells and CEFs infected with recombinant vaccinia virus. (A) Schematic diagram of the genetic organization of the pJSA1175 DNA vector (top). The bottom shows the cDNA fragments of influenza virus BJ95 (H3N2) PB1, NP, and M1 after restriction digestion with BamHI or SmaI, as appropriate, and smoothing, respectively. (B) Indirect immunofluorescence showing the expression of influenza PB1, NP, and M1 in plaques formed in rVV-PB1 (left)-, rVV-NP (middle)-, and rVV-M1 (right)-infected HeLa cells (upper) and stained with the polyclonal or monoclonal Abs indicated in Materials and Methods. The bottom images show the results of HeLa cells mock infected with rVV and analyzed using the same Abs as in the top images. (C) Western blots showing the expression of influenza PB1, NP, and M1 from rVV-PB1 (lane 1)-, rVV-NP (lane 2)-, and rVV-M1 (lane 3)-infected CEF cell lysates stained with the polyclonal or monoclonal antiserum indicated in Materials and Methods. M, molecular weight marker; Neg, cell lysates from CEFs mock infected with rVV-c.