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Fig. 1 | Virology Journal

Fig. 1

From: An equine herpesvirus type 1 (EHV-1) vector expressing Rift Valley fever virus (RVFV) Gn and Gc induces neutralizing antibodies in sheep

Fig. 1

Generation of recombinant EHV-1 expressing Gn-Gc protein of RVFV (rH_Gn-Gc). Schematic illustration of the construction of rH_Gn-Gc vaccine vector based on pRacH1. a Depiction of the left terminus of the unique-long segment of EHV-1 strain RacH infectious BAC clone pH 1-EF1, in which ORF1 and ORF2 are naturally deleted. b A fragment released from transfer plasmid pUC19-ORF1/2-Gn-Gc by I-CeuI digestion was used to recombine with RacH genome, result in incorporation of Gn-Gc gene of RVFV, HCMV promoter and kanamycin resistance gene in the ORF1/ORF2 locus of the RacH genome. c After I-SceI digestion, kanamycin was removed in the following step of en passent mutagenesis to generate the final arrangement of rH_Gn-Gc genome. d and e Restriction fragment length polymorphisms and southern blot of pH1_EF1, the cloning intermediate and the final pH_Gn-Gc construct. An ethidium bromide-stained agarose gel is shown in the left panel with EcoRV restriction patterns of pH_EF1 (lanes 1), the kanamycin-resistant intermediate (lanes 2) and pH_Gn-Gc (lanes 3). GeneRuler 1 kb Plus DNA Ladder (Thermo Scientific) was used for determination of DNA fragment sizes. In the right panel, a southern blot of the same gel is shown after hybridization with a digoxigenin-labeled Gn-Gc RVFV probe

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