Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F05%3A00026599" target="_blank" >RIV/61388971:_____/05:00026599 - isvavai.cz</a>
Result on the web
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DOI - Digital Object Identifier
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Alternative languages
Result language
angličtina
Original language name
Post-Translational Modification(s) and Cell Distribution of Streptomyces aureofaciens Translation Elongation Factor Tu Overproduced in Escherichia coli
Original language description
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 bodiesformed 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
Czech name
Post-translační modifikace a distribuce translačního elongačního faktoru Tu ze Streptomyces aureofaciens nadprodukovaného v Escherichia coli
Czech description
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
Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EE - Microbiology, virology
OECD FORD branch
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Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
Z - Vyzkumny zamer (s odkazem do CEZ)
Others
Publication year
2005
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Folia Microbiologica
ISSN
0015-5632
e-ISSN
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Volume of the periodical
50
Issue of the periodical within the volume
5
Country of publishing house
CZ - CZECH REPUBLIC
Number of pages
8
Pages from-to
393-400
UT code for WoS article
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EID of the result in the Scopus database
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