Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/32836
DC FieldValueLanguage
dc.contributor.authorLin, H.W.en_US
dc.contributor.author張天傑zh_TW
dc.contributor.authorChang, Y.Y.en_US
dc.contributor.authorWong, M.L.en_US
dc.contributor.authorLin, J.W.en_US
dc.contributor.authorChang, T.J.en_US
dc.date2004zh_TW
dc.date.accessioned2014-06-06T07:44:16Z-
dc.date.available2014-06-06T07:44:16Z-
dc.identifier.issn0042-6822zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/32836-
dc.description.abstractDuring lytic infection, the virion host shutoff (vhs) protein of alphaherpesviruses causes the degradation of mRNAs nonspecifically. In this work, we cloned the vhs gene (UL41 open reading frame) of pseudorabies virus (PRV; TNL strain) by PCR, and its nucleotide sequences were determined. The PCR product of vhs gene was subcloned into the prokaryotic pET32b expression vector, and production of the recombinant vhs protein was examined by SDS-PAGE. Result of Western blotting demonstrated that our recombinant vhs protein reacted with antiserum against a synthetic peptide of 17 amino acids of the vhs protein. After purification with nickel-chelate affinity chromatography, the purified recombinant vhs protein exhibited in vitro ribonuclease activity as expected. We further cloned the vhs gene into eukaryotic expression vectors and investigated the intracellular function of vhs protein by DNA transfection. By transient transfection and CAT assay, we found the CAT activity was reduced in the presence of vhs, indicating that degradation of mRNA of the CAT gene was caused by the vhs. Furthermore, our results showed that the plaque formation of pseudorabies virus was blocked by exogenous vhs. Taken together, we have cloned the vhs gene of pseudorabies virus (TNL strain) and conducted functional analysis of the recombinant vhs protein in vitro as well as in vivo. (C) 2004 Elsevier Inc. All rights reserved.en_US
dc.language.isoen_USzh_TW
dc.relationVirologyen_US
dc.relation.ispartofseriesVirology, Volume 324, Issue 2, Page(s) 412-418.en_US
dc.relation.urihttp://dx.doi.org/10.1016/j.virol.2004.04.015en_US
dc.subjectpseudorabies virusen_US
dc.subjectvhs geneen_US
dc.subjectherpes-simplex-virusen_US
dc.subjectmessenger-rna degradationen_US
dc.subjectdna-sequenceen_US
dc.subjectequineen_US
dc.subjectherpesvirus-1en_US
dc.subjectvhs proteinen_US
dc.subjectgene ul41en_US
dc.subjectin-vitroen_US
dc.subjecttype-1en_US
dc.subjectcellsen_US
dc.subjectidentificationen_US
dc.titleFunctional analysis of virion host shutoff protein of pseudorabies virusen_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1016/j.virol.2004.04.015zh_TW
item.languageiso639-1en_US-
item.openairetypeJournal Article-
item.cerifentitytypePublications-
item.grantfulltextnone-
item.fulltextno fulltext-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
Appears in Collections:獸醫學系所
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