表达羊口疮病毒B2L蛋白重组小反刍兽疫病毒的拯救

Rescue of Recombinant Peste des Petits Ruminants Virus for Expressing Orf Virus B2L Protein

羊口疮(传染性脓疱)和小反刍兽疫的病原体分别为羊口疮病毒(orf virus,ORFV)和小反刍兽疫病毒(peste des petits ruminants virus,PPRV)。如果通过反向遗传技术进行基因修饰,PPRV可成为表达外源蛋白的有效载体。ORFV含有1个具有高免疫原性的B2L蛋白。本研究首先构建了含B2L基因开放阅读框的重组PPRV cDNA clone,然后通过反向遗传技术,拯救了一株表达B2L蛋白的重组PPRV。试验表明,该重组病毒的生长动力学类似其母源病毒;Western blot和质谱分析表明,该重组PPRV可在细胞内表达B2L蛋白。该研究为羊口疮和小反刍兽疫二联疫苗的研制奠定了基础。

Orf(contagious ecthyma)and peste des petits ruminants(PPR)are caused by orf virus(ORFV)and peste des petits ruminants virus(PPRV),respectively. If genetically modified by reverse genetics tool,the PPRV would be a useful vector to express foreign proteins. The ORFV contains a highly immunogenic envelope protein,B2L protein. In this study,a recombinant PPRV cDNA clone,which contained the B2L gene open reading frame,was rescued using reverse genetics to express the B2L protein. This recombinant PPRV was demonstrated to have similar growth kinetics to that of its parental virus. In vitro expression of the B2L protein by it was demonstrated by Western blot and confirmed by mass spectrometry,therefore paving the way for development of a bivalent vaccine candidate against both diseases.