About: Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli     Goto   Sponge   NotDistinct   Permalink

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  • We cloned EF-Tu from Streptomyces aureofaciens on a pET plasmid and overproduced it using the T7 RNA polymerase system in Escherichia coli. Streptomyces EF-Tu represented more than 40 % of the total cell protein and was stored mostly in inclusion bodies formed apically at both ends of E. coli cells. Analysis of the inclusion bodies by transmission and scanning electron microscopy did not reveal any inter-nal or surface ultrastructures. We developed the method for purification of S. aureofaciens EF-Tu from iso-lated inclusion bodies based on the ability of the protein to aggregate spontaneously. EF-Tu present in inclusion bodies was not active in GDP binding. Purified protein showed a similar charge heterogeneity as EF-Tu isolated from the mycelium of S. aureofaciens and all of the isoforms reacted with EF-Tu antibodies. All isoforms also reacted with monoclonal antibodies against O-phosphoserine and O-phosphothreonine
  • We cloned EF-Tu from Streptomyces aureofaciens on a pET plasmid and overproduced it using the T7 RNA polymerase system in Escherichia coli. Streptomyces EF-Tu represented more than 40 % of the total cell protein and was stored mostly in inclusion bodies formed apically at both ends of E. coli cells. Analysis of the inclusion bodies by transmission and scanning electron microscopy did not reveal any inter-nal or surface ultrastructures. We developed the method for purification of S. aureofaciens EF-Tu from iso-lated inclusion bodies based on the ability of the protein to aggregate spontaneously. EF-Tu present in inclusion bodies was not active in GDP binding. Purified protein showed a similar charge heterogeneity as EF-Tu isolated from the mycelium of S. aureofaciens and all of the isoforms reacted with EF-Tu antibodies. All isoforms also reacted with monoclonal antibodies against O-phosphoserine and O-phosphothreonine (en)
  • Klonovali jsme EF-Tu z S. aureofaciens na plasmidu pET a nadprodukovali ho v E. coli. Streptomycetový EF-Tu představoval více než 40% všech bílkovin a byl uložen převážně v inkluzích. Analýza inkluzí neodhalila žádné vnitřní ani povrchové ultrastruktury. Vyvinuli jsme metodu purifikace faktoru z inkluzí. Purifikovaný protein vykazoval stejnou nábojovou heterogenitu jako faktor ve streptomycetách. Všechny izoformy reagovaly s monoklonálními protilátkami proti O-phosphoserinu a O-phosphothreoninu (cs)
Title
  • Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli
  • Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli (en)
  • Post-translační modifikace a distribuce translačního elongačního faktoru Tu ze Streptomyces aureofaciens nadprodukovaného v Escherichia coli (cs)
skos:prefLabel
  • Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli
  • Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli (en)
  • Post-translační modifikace a distribuce translačního elongačního faktoru Tu ze Streptomyces aureofaciens nadprodukovaného v Escherichia coli (cs)
skos:notation
  • RIV/61388971:_____/05:00026599!RIV06-AV0-61388971
http://linked.open.../vavai/riv/strany
  • 393;400
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(GA204/00/1252), P(GA204/03/1011), Z(AV0Z50200510)
http://linked.open...iv/cisloPeriodika
  • 5
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 537175
http://linked.open...ai/riv/idVysledku
  • RIV/61388971:_____/05:00026599
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • streptomyces aureofaciens; Escherichia coli; post-translational modification (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • CZ - Česká republika
http://linked.open...ontrolniKodProRIV
  • [A1025C8B02C7]
http://linked.open...i/riv/nazevZdroje
  • Folia Microbiologica
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 50
http://linked.open...iv/tvurceVysledku
  • Benada, Oldřich
  • Weiserová, Marie
  • Weiser, Jaroslav
  • Kofroňová, Olga
  • Holub, Martin
  • Kalachová, Ladislava
  • Nguyen, Liem Duy
  • Kormanec, J.
http://linked.open...n/vavai/riv/zamer
issn
  • 0015-5632
number of pages
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