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. 2015 Jun 18;9(6):e0003850. doi: 10.1371/journal.pntd.0003850

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© 2015 Franceschi et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 2 — Diagram (not to scale; A) showing the retargeting event obtained by heat-inducible homologous recombination in SW102 containing pBAC-BoHV-4-A-TK-KanaGalK-TK, where the Kana/GalK cassette was replaced with the CMV-A29LgD₁₀₆, EF1α-M1RgD₁₀₆ and EF1α-B6RgD₁₀₆ expression cassettes flanked by BoHV-4 TK sequences, located in pINT2 shuttle plasmid vector. B) Representative 2-deoxy-galactose resistant colonies (1 and 2) tested by HindIII restriction enzyme analysis, agar gel electrophoresis, and Southern blotting performed with specific probes for the A29L, M1R and B6R ORFs. The 2,650 bp band (yellow circle), corresponding to the un-retargeted pBAC-BoHV-4-A-TK-KanaGalK-TK control (C), has been replaced by a 1,661 bpband (red circle) in pBAC-BoHV-4-A-CMV-A29LgD₁₀₆ΔTK, by a 2,929 bp band (red circle) in pBAC-BoHV-4-A-EF1α-B6RgD₁₀₆ΔTK and by a 2,725 bp band (red circle) in pBAC-BoHV-4-A-EF1α-M1RgD₁₀₆ΔTK.