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Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli
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SYSNO ASEP 0026599 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Ostatní články Title Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli Title Post-translační modifikace a distribuce translačního elongačního faktoru Tu ze Streptomyces aureofaciens nadprodukovaného v Escherichia coli Author(s) Nguyen, Liem Duy (MBU-M)
Holub, Martin (MBU-M)
Kalachová, Ladislava (MBU-M)
Weiserová, Marie (MBU-M) RID
Kormanec, J. (SK)
Benada, Oldřich (MBU-M) ORCID, RID
Kofroňová, Olga (MBU-M) RID, ORCID
Weiser, Jaroslav (MBU-M) RIDSource Title Folia Microbiologica. - : Springer - ISSN 0015-5632
Roč. 50, č. 5 (2005), s. 393-400Number of pages 8 s. Language eng - English Country CZ - Czech Republic Keywords streptomyces aureofaciens ; Escherichia coli ; post-translational modification Subject RIV EE - Microbiology, Virology R&D Projects GA204/00/1252 GA ČR - Czech Science Foundation (CSF) GA204/03/1011 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z50200510 - MBU-M (2005-2011) Annotation 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 Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2006
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