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. 2013 Nov 5;8(11):e78834. doi: 10.1371/journal.pone.0078834

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© 2013 Simón 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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A) Donor plasmids used in this study. B) Viruses used in this study. The sequence of the complete genotype SfNIC-B is shown, where the right and left pif-2/pif-1 flanking regions are specified. First, a SfNIC-BΔpifs recombinant virus was constructed in which pif-2/pif-1 coding region was substituted by the LacZ operon. The primers used for the flanking regions amplifications are indicated below the figure. Recombinant viruses SfNIC-Bpif1 (rescue virus), SfNIC-Begt, in which the SeMNPV egt promoter would drive pif-1 transcription, and SfNIC-Bp10, in which the SeMNPV p10 promoter would drive pif-1 transcription, were constructed by homologous recombination between SfNIC-BΔpifs and donor plasmids pUC19.(pif1)pif1, pUC19.(egt)pif1 and pUC19.(p10)pif1 respectively. C) pif-2/pif-1 intergenic sequences from SfNIC-B, SfNIC-Bpif1, SfNIC-Begt and SfNIC-Bp10 viruses. The underlined portion in SfNIC-B containing the native pif-1 promoter was replaced by donor sequences.