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微生物学报 2006
Co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus can form heterodimers
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Abstract:
In order to investigate the characterization of in vitro co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus(PRRSV),eukaryotic expression plasmids pCI-ORF5(expressing GP5 protein alone),pCI-ORF6(expressing M protein alone),and pCI-ORF5/ORF6(co-expressing GP5 and M proteins) were constructed.After transient transfection,Western blot analysis under nonreducing condition demonstrated that co-expressed GP5 and M proteins could form disulfide-linked heterodimers(GP5-M) in transiently transfected BHK-21 cells.To further study the influence of GP5-M heterodimers formation on the subcellular localizations of GP5 or M proteins,green fluorescence protein(EGFP) and red fluorescence protein(RFP) were used as markers.The results of fluorescence distribution showed that co-expressed GP5-EGFP chimera and M-RFP chimera boosted the transport of GP5 from the endoplasmic reticulum(ER) to the Golgi complex,indicating that the formation of GP5-M heterodimers may be involved in posttranslational modification,transport,and subcellular localization of GP5.These results presented here lay foundation to further study the molecular mechanism of GP5-M heterodimer formation and its role in protective immunity of PRRSV.