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. 2010 Nov 10;85(2):1011–1024. doi: 10.1128/JVI.01092-10

FIG. 2.

FIG. 2.

Production of a recombinant BoHV-4 strain deleted for Bo10. Using the BoHV-4 V. test strain as the parental strain, a recombinant BoHV-4 strain deleted for Bo10 and a derived revertant strain were produced by homologous recombination. (A) Recombination cassettes were constructed. The HindIII restriction map of the entire BoHV-4 V. test strain is shown at the top. Letters were assigned to the different restriction fragments according to their lengths, as described by Bublot et al. (4a). Most of Bo10 exons 1 and 2 were replaced by an eGFP expression cassette as described in Materials and Methods. prDNA, polyrepetitive DNA; CMV IE promoter, cytomegalovirus immediate-early promoter; SV40 pA, simian virus 40 pA. (B) Bo10 Del and Bo10 Rev strains were then characterized by a combined restriction endonuclease and Southern blotting approach. The DNA of the parental strain V. test and the derived recombinant strains Bo10 Del and Bo10 Rev were analyzed by HindIII restriction and further tested by Southern blotting using probes corresponding to the entire V. test Bo10 ORF or to the eGFP ORF or to the deleted part of the V. test Bo10 ORF. The asterisk and arrows indicate the restriction fragments containing the entire Bo10 and eGFP ORFs, respectively. Marker sizes (in kilobase pairs) are indicated on the left.