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Article: Identification of O-antigen polymerase transcription and translation start signals and visualization of the protein in Salmonella enterica serovar Typhimurium

TitleIdentification of O-antigen polymerase transcription and translation start signals and visualization of the protein in Salmonella enterica serovar Typhimurium
Authors
Issue Date1999
PublisherSociety for General Microbiology. The Journal's web site is located at http://mic.sgmjournals.org
Citation
Microbiology, 1999, v. 145 n. 9, p. 2443-2451 How to Cite?
AbstractThe wzy/rfe gene, encoding the O-antigen polymerase, of Salmonella enterica serovar Typhimurium has been previously cloned and sequenced. In the present work, the wry transcriptional startpoint was initially identified by primer extension. Next, wzy promoter strength in Escherichia coli K-12 was measured, and was found to be greater than that of the induced lac promoter. To define the Wzy translational startpoint, DNA including the wry promoter and the putative first five residues of the Wzy protein was fused to the N-terminus of glutathione-S-transferase, and the fusion protein purified by affinity chromatography. N-terminal amino acid sequencing yielded the Wzy translational startpoint. Next, the Wzy protein was C-terminally tagged with the FLAG peptide, and immunoblotting of an S. typhimurium strain expressing a low-copy wzy-FLAG gene (five copies per cell) localized the intact Wzy protein in the cytoplasmic membrane of S. typhimurium cells. The Wzy protein was not well-expressed from a multi-copy wzy-FLAG + plasmid in S. typhimurium, or in E. coli K-12.
Persistent Identifierhttp://hdl.handle.net/10722/162310
ISSN
2015 Impact Factor: 2.268
2015 SCImago Journal Rankings: 1.144
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWong, DKHen_US
dc.contributor.authorMorris, Cen_US
dc.contributor.authorLam, TLen_US
dc.contributor.authorWong, WKRen_US
dc.contributor.authorHackett, Jen_US
dc.date.accessioned2012-09-05T05:18:52Z-
dc.date.available2012-09-05T05:18:52Z-
dc.date.issued1999en_US
dc.identifier.citationMicrobiology, 1999, v. 145 n. 9, p. 2443-2451en_US
dc.identifier.issn1350-0872en_US
dc.identifier.urihttp://hdl.handle.net/10722/162310-
dc.description.abstractThe wzy/rfe gene, encoding the O-antigen polymerase, of Salmonella enterica serovar Typhimurium has been previously cloned and sequenced. In the present work, the wry transcriptional startpoint was initially identified by primer extension. Next, wzy promoter strength in Escherichia coli K-12 was measured, and was found to be greater than that of the induced lac promoter. To define the Wzy translational startpoint, DNA including the wry promoter and the putative first five residues of the Wzy protein was fused to the N-terminus of glutathione-S-transferase, and the fusion protein purified by affinity chromatography. N-terminal amino acid sequencing yielded the Wzy translational startpoint. Next, the Wzy protein was C-terminally tagged with the FLAG peptide, and immunoblotting of an S. typhimurium strain expressing a low-copy wzy-FLAG gene (five copies per cell) localized the intact Wzy protein in the cytoplasmic membrane of S. typhimurium cells. The Wzy protein was not well-expressed from a multi-copy wzy-FLAG + plasmid in S. typhimurium, or in E. coli K-12.en_US
dc.languageengen_US
dc.publisherSociety for General Microbiology. The Journal's web site is located at http://mic.sgmjournals.orgen_US
dc.relation.ispartofMicrobiologyen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCell Fractionationen_US
dc.subject.meshCulture Mediaen_US
dc.subject.meshElectrophoresis, Polyacrylamide Gelen_US
dc.subject.meshEnzyme-Linked Immunosorbent Assayen_US
dc.subject.meshGenes, Bacterialen_US
dc.subject.meshHexosyltransferases - Analysis - Chemistry - Geneticsen_US
dc.subject.meshImmunoblottingen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPeptides - Analysisen_US
dc.subject.meshPromoter Regions, Geneticen_US
dc.subject.meshProtein Biosynthesisen_US
dc.subject.meshRecombinant Fusion Proteins - Analysis - Chemistry - Isolation & Purificationen_US
dc.subject.meshSalmonella Typhimurium - Enzymology - Genetics - Growth & Developmenten_US
dc.subject.meshTranscription, Geneticen_US
dc.titleIdentification of O-antigen polymerase transcription and translation start signals and visualization of the protein in Salmonella enterica serovar Typhimuriumen_US
dc.typeArticleen_US
dc.identifier.emailWong, DKH:danywong@hku.hken_US
dc.identifier.authorityWong, DKH=rp00492en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid10517597-
dc.identifier.scopuseid_2-s2.0-0032879212en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032879212&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume145en_US
dc.identifier.issue9en_US
dc.identifier.spage2443en_US
dc.identifier.epage2451en_US
dc.identifier.isiWOS:000082829000027-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridWong, DKH=7401535819en_US
dc.identifier.scopusauthoridMorris, C=7401472993en_US
dc.identifier.scopusauthoridLam, TL=55046412000en_US
dc.identifier.scopusauthoridWong, WKR=7403972071en_US
dc.identifier.scopusauthoridHackett, J=24302040000en_US

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