羊口瘡(傳染性膿皰)和小反芻獸疫的病原體分別為羊口瘡病毒(orf virus,,ORFV)和小反芻獸疫病毒(peste despetits ruminants virus,,PPRV)。如果通過反向遺傳技術(shù)進(jìn)行基因修飾,,PPRV可成為表達(dá)外源蛋白的有效載體,。ORFV含有1個(gè)具有高免疫原性的B2L蛋白。本研究首先構(gòu)建了含B2L基因開放閱讀框的重組PPRV cDNA clone,,然后通過反向遺傳技術(shù),,拯救了一株表達(dá)B2L蛋白的重組PPRV。試驗(yàn)表明,該重組病毒的生長(zhǎng)動(dòng)力學(xué)類似其母源病毒,;Western blot和質(zhì)譜分析表明,,該重組PPRV可在細(xì)胞內(nèi)表達(dá)B2L蛋白。該研究為羊口瘡和小反芻獸疫二聯(lián)疫苗的研制奠定了基礎(chǔ),。
Rescue of Recombinant Peste des PetitsRuminants Virus for Expressing Orf Virus B2L Protein
Orf(contagiousecthyma)and peste des petits ruminants(PPR)are caused byorf virus(ORFV)and peste despetits ruminants virus(PPRV),,respectively. If genetically modifiedby reverse genetics tool,the PPRV wouldbe a useful vector to express foreign proteins. The ORFV contains a highlyimmunogenic envelope protein,,B2L protein.In this study,,a recombinant PPRV cDNA clone,which contained the B2L gene openreading frame,,was rescued using reverse genetics toexpress the B2L protein. This recombinant PPRV was demonstrated to have similargrowth kinetics to that of its parental virus. In vitro expression of the B2Lprotein by it was demonstrated by Western blot and confirmed by massspectrometry,,therefore paving the way fordevelopment of a bivalent vaccine candidate against both diseases.
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