Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/71573
DC FieldValueLanguage
dc.contributor.authorYin, H.S.en_US
dc.contributor.authorShien, J.H.en_US
dc.contributor.authorLee, L.H.en_US
dc.date2000zh_TW
dc.date.accessioned2014-06-11T06:01:59Z-
dc.date.available2014-06-11T06:01:59Z-
dc.identifier.issn0042-6822zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/71573-
dc.description.abstractThe genome segment S2 of avian reovirus (ARV) S1133 was cloned and sequenced. The entire S2 nucleotide sequence is 1325 bp long with one long open reading frame that encodes a protein of 415 amino acids, corresponding to sigma A, a major core protein of ARV. S2 possesses a pentanucleotide, TCATC, at the 3'-terminus of its plus strand, common to other known genome segments of ARV and to 10 genome segments of mammalian reovirus. Amino acid sequence analysis revealed that sigma A contains a carboxy-terminal region (one-fourth of the protein) mat is formed from alpha-helices and beta-turns, and the remainder (three-fourths of the protein) is formed predominantly from beta-strands and beta-turns. Analysis of binding activity to poly(rl)-poly(rC)-agarose suggested that ARV protein A present in total virus-infected chicken embryo fibroblasts (CEF) had dsRNA-binding activity. To further characterize the binding activity, protein sigma A was subsequently expressed in Escherichia coli BL21(DE3) cells as a fusion protein and isolated by metal chef ate affinity chromatography. The expressed protein e sigma A was further purified through a Superdex 75 HR 10/30 column after digestion of me purified fusion peptide with enterokinase. The expressed protein e sigma A has the same molecular weight as virion protein sigma A purified from ARV-infected CEF and is indistinguishable from virion protein sigma A by immunoblot analysis. The e sigma A binds cooperatively alpha P-32-labeled dsRNA probe produced by run-off transcription of clone pGEM-3Zf(+)S4. The binding reaction is blocked by homologous ARV dsRNA or heterologous infectious bursal disease virus dsRNA and poly(rl)-poly(rC), but not by salmon sperm DNA. The results indicate that the expressed protein e sigma A has dsRNA-binding activity similar to that of sigma A obtained from infected cells, and its binding is sequence-independent. (C) 2000 Academic Press.en_US
dc.language.isoen_USzh_TW
dc.relationVirologyen_US
dc.relation.ispartofseriesVirology, Volume 266, Issue 1, Page(s) 33-41.en_US
dc.relation.urihttp://dx.doi.org/10.1006/viro.1999.0020en_US
dc.subjectdouble-stranded-rnaen_US
dc.subjectstructural proteinsen_US
dc.subjectcoding assignmenten_US
dc.subjectgenomeen_US
dc.subjectsegmentsen_US
dc.subjectstrain s1133en_US
dc.subjectvirusen_US
dc.subjectgeneen_US
dc.subjectpolypeptidesen_US
dc.subjectsequencesen_US
dc.subjectcloningen_US
dc.titleSynthesis in Escherichia coli of avian reovirus core protein sigma A and its dsRNA-binding activityen_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1006/viro.1999.0020zh_TW
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeJournal Article-
item.cerifentitytypePublications-
item.fulltextno fulltext-
item.languageiso639-1en_US-
item.grantfulltextnone-
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