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2011-09-24Zeitschriftenartikel DOI: 10.1016/j.pep.2011.09.006
Optimisation of expression and purification of the feline and primate foamy virus transmembrane envelope proteins using a 96 deep well screen
dc.contributor.authorMühle, Michael
dc.contributor.authorLöchelt, Martin
dc.contributor.authorDenner, Joachim
dc.date.accessioned2018-05-07T15:56:39Z
dc.date.available2018-05-07T15:56:39Z
dc.date.created2012-09-24
dc.date.issued2011-09-24none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/rewlNXXdroqzk/PDF/29ZkPaEBMb4IQ.pdf
dc.identifier.urihttp://edoc.rki.de/176904/1313
dc.description.abstractThe production of recombinant transmembrane proteins is due to their biochemical properties often troublesome and time consuming. Here the prokaryotic expression and purification of the transmembrane envelope proteins of the feline and primate foamy viruses using a screening assay for optimisation of expression in 96 deep well plates is described. Testing simultaneously various bacterial strains, media, temperatures, inducer concentrations and different transformants, conditions for an about twentyfold increased production were quickly determined. These small scale test conditions could be easily scaled up, allowing purification of milligram amounts of recombinant protein. Proteins with a purity of about 95% were produced using a new purification protocol, they were characterised by gel filtration and circular dichroism and successfully applied in immunological assays screening for foamy virus infection and in immunisation studies. Compared to the previously described protocol (M. Mühle, A. Bleiholder, S. Kolb, J. Hübner, M. Löchelt, J. Denner, Immunological properties of the transmembrane envelope protein of the feline foamy virus and its use for serological screening, Virology 412 (2011) 333–340), proteins with similar characteristics but about thirtyfold increased yields were obtained. The screening and production method presented here can also be applied for the production of transmembrane envelope proteins of other retroviruses, including HIV-1.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionskrankheiten / Erreger
dc.subjectTransmembrane envelope proteineng
dc.subjectOptimisation of protein expressioneng
dc.subjectHigh throughput screeningeng
dc.subjectFoamy viruseseng
dc.subjectHIV-1eng
dc.subject.ddc610 Medizin
dc.titleOptimisation of expression and purification of the feline and primate foamy virus transmembrane envelope proteins using a 96 deep well screen
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10027307
dc.identifier.doi10.1016/j.pep.2011.09.006
dc.identifier.doihttp://dx.doi.org/10.25646/1238
local.edoc.container-titleProtein Expression and Purification
local.edoc.container-textMühle, M., Löchelt, M., Denner, J. Optimisation of expression and purification of the feline and primate foamy virus transmembrane envelope proteins using a 96 deep well screen (2012) Protein Expression and Purification, 81 (1), pp. 96-105.
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.sciencedirect.com/science/article/pii/S1046592811002531
local.edoc.container-publisher-nameElsevier
local.edoc.container-volume81
local.edoc.container-issue1
local.edoc.container-year2012

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