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Figure 1 | Veterinary Research

Figure 1

From: Evaluation of immune responses following infection of ponies with an EHV-1 ORF1/2 deletion mutant

Figure 1

(A). Genomic organization of RacH, Ab4 wild type and the recombinant Ab4 OFR1/2 deletion mutant. Shown is the RacH and Ab4genome with a detailed organization of parts of the unique long (UL) and unique short (US) regions, along with parts of the inverted and terminal repeat regions (IR & TR, shaded in grey). In addition, the genome of the recombinant Ab4 ORF1/2 mutant is shown where the genes ORF1 and ORF2 were deleted. B. Restriction fragment length polymorphisms (RFLPs) that confirm correct deletion of the ORF1 and 2 genes in the pAb4 background. BAC DNA was digested with NotI and separated by electrophoresis on a 0.8% agarose gel. Staining of the gel with ethidium bromide (EtBr) shows changes in RFLP patters obtained, indicated by circles, after the first recombination (insertion of the aphA1 gene; pAb4 intermediate) and the second recombination (removal of aphA1 gene and the ORF1/2 genes; pAb4ΔORF1/2). (C). Western blot analysis detecting the expression of gp2 in cellysates of cells infected with reconstituted or not reconstituted Ab4ΔORF1/2. Mock-infected cells (RK13 cells) were included as controls. Cellysates were loaded and separated by SDS-12% PAGE under reducing conditions. Separated proteins were transferred to a nitrocellulose membrane and incubated with monoclonal antibodies (mAb) 3B12 (against gp2; 1:10) or the control mAb L3ab (against ETIF, 1:2000). The sizes of the PageRuler Prestained protein Ladder (Fermentas) are given in thousands.

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