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. 2009 Jun 18;40(5):51. doi: 10.1051/vetres/2009034

Figure 1.

Figure 1.

Production and characterization of BoHV-4 recombinant strains expressing membrane anchored recombinant forms of IRAC I and IRAC II. (A) IRAC I CD46, -I HA, -II CD46 or -II HA expression cassette were inserted in the BstEII site of the V. test BAC G plasmid corresponding to the far left BstEII site of BoHV-4 L-DNA. The insertion was performed by mutagenesis of the V. test BAC G plasmid in bacteria through homologous recombination with pST76KSR-lacZ IRAC I HA, -I CD46, -II HA or -II CD46 plasmids. (B and C) Characterization of BoHV-4 recombinant strains expressing membrane anchored IRAC by a combined restriction endonuclease-Southern blot approach. The DNA of the modified BAC plasmids generated in bacteria or from the reconstituted viral strains were analysed by HindIII restriction (B and C, left panels) and further tested by Southern blot using IRAC I ORF (B, middle panel), IRAC II ORF (B, right panel), human CD46 ORF (C, middle panel) and Vaccinia HA ORF (C, right panel) as probes. Arrows indicate the restriction fragments containing IRAC ORF. Marker sizes (MS) in kb are indicated on the left.