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2005-06-20Zeitschriftenartikel DOI: 10.1186/1472-6750-5-19
Viral promoters can initiate expression of toxin genes introduced into Escherichia coli
dc.contributor.authorLewin, Astrid
dc.contributor.authorMayer, Martin
dc.contributor.authorChusainow, Janet
dc.contributor.authorJacob, Daniela
dc.contributor.authorAppel, Bernd
dc.date.accessioned2018-05-07T13:07:25Z
dc.date.available2018-05-07T13:07:25Z
dc.date.created2009-04-24
dc.date.issued2005-06-20none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/re3QNNiiUpr5k/PDF/270dJYfBVkzl.pdf
dc.identifier.urihttp://edoc.rki.de/176904/395
dc.description.abstractBackground: The expression of recombinant proteins in eukaryotic cells requires the fusion of the coding region to a promoter functional in the eukaryotic cell line. Viral promoters are very often used for this purpose. The preceding cloning procedures are usually performed in Escherichia coli and it is therefore of interest if the foreign promoter results in an expression of the gene in bacteria. In the case molecules toxic for humans are to be expressed, this knowledge is indispensable for the specification of safety measures. Results: We selected five frequently used viral promoters and quantified their activity in E. coli with a reporter system. Only the promoter from the thymidine kinase gene from HSV1 showed no activity, while the polyhedrin promoter from baculovirus, the early immediate CMV promoter, the early SV40 promoter and the 5' LTR promoter from HIV-1 directed gene expression in E. coli. The determination of transcription start sites in the immediate early CMV promoter and the polyhedrin promoter confirmed the existence of bacterial -10 and -35 consensus sequences. The importance of this heterologous gene expression for safety considerations was further supported by analysing fusions between the aforementioned promoters and a promoter-less cytotoxin gene. Conclusion: According to our results a high percentage of viral promoters have the ability of initiating gene expression in E. coli. The degree of such heterologous gene expression can be sufficient for the expression of toxin genes and must therefore be considered when defining safety measures for the handling of corresponding genetically modified organisms.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionskrankheiten / Erreger
dc.subjectHumanseng
dc.subjectEscherichia coli/geneticseng
dc.subjectCloningeng
dc.subjectMoleculareng
dc.subjectMolecular Sequence Dataeng
dc.subjectBacterial Proteins/geneticseng
dc.subjectBaculoviridae/geneticseng
dc.subjectBase Sequenceeng
dc.subjectBiotechnology/methodseng
dc.subjectCytomegalovirus/geneticseng
dc.subjectCytotoxins/geneticseng
dc.subjectDNA/analysiseng
dc.subjectEscherichia coli/metabolismeng
dc.subjectGene Expression Regulationeng
dc.subjectBacterialeng
dc.subjectViraleng
dc.subjectGeneseng
dc.subjectReportereng
dc.subjectHIV-1/geneticseng
dc.subjectHemolysin Proteins/metabolismeng
dc.subjectHemolysiseng
dc.subjectLuciferases/metabolismeng
dc.subjectLuminescenceeng
dc.subjectModelseng
dc.subjectGeneticeng
dc.subjectPlasmids/metabolismeng
dc.subjectPromoter Regionseng
dc.subjectSimian virus 40/geneticseng
dc.subjectTerminal Repeat Sequenceseng
dc.subjectThymidine Kinase/geneticseng
dc.subjectThymidine Kinase/metabolismeng
dc.subjectToxinseng
dc.subjectBiological/metabolismeng
dc.subjectTranscription Initiation Siteeng
dc.subjectTranscriptioneng
dc.subjectViral Proteins/geneticseng
dc.subjectViral Structural Proteinseng
dc.subject.ddc610 Medizin
dc.titleViral promoters can initiate expression of toxin genes introduced into Escherichia coli
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-100331
dc.identifier.doi10.1186/1472-6750-5-19
dc.identifier.doihttp://dx.doi.org/10.25646/320
local.edoc.container-titleBMC Biotechnology
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.biomedcentral.com/1472-6750/5/19
local.edoc.container-publisher-nameBioMed Central
local.edoc.container-volume5
local.edoc.container-issue19
local.edoc.container-year2005

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