Linked Life Data

8535157

Source:http://linkedlifedata.com/resource/pubmed/id/8535157

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
1996-2-8
pubmed:abstractText
The translation elongation factors G (EF-G), Tu (EF-Tu), and Ts (EF-Ts) from the extreme thermophilic bacterium Thermus thermophilus were overproduced in Escherichia coli. The fus gene coding for EF-G and the tufA gene coding for EF-Tu were expressed under the control of a tac promoter, whereas EF-Ts was overproduced with the T7 RNA polymerase system. A detailed description for the purification of the three elongation factors from E. coli is presented. EF-G and EF-Tu are isolated by Q-Sepharose FF chromatography, heat treatment at 65 or 60 degrees C, respectively, and Sephacryl S200 gel permeation chromatography. For the purification of EF-Ts, a heat denaturation step is followed by DEAE-cellulose chromatography and a cation exchange EMD-SO-3 650 column. The overproduced factors show the same properties as those purified from T. thermophilus. As the crystal structures of T. thermophilus EF-Tu and EF-G have been solved recently, many questions concerning the function of particular residues or domains arise, which may be best addressed by studying the in vitro behavior and structure of altered recombinant constructs. The methods presented here should facilitate such studies.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
1046-5928
pubmed:author
pubmed:issnType
Print
pubmed:volume
6
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
637-45
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:8535157-Bacteriophage T7, pubmed-meshheading:8535157-Base Sequence, pubmed-meshheading:8535157-Centrifugation, pubmed-meshheading:8535157-Chromatography, Ion Exchange, pubmed-meshheading:8535157-Cloning, Molecular, pubmed-meshheading:8535157-Escherichia coli, pubmed-meshheading:8535157-Hot Temperature, pubmed-meshheading:8535157-Molecular Sequence Data, pubmed-meshheading:8535157-Muramidase, pubmed-meshheading:8535157-Peptide Elongation Factor G, pubmed-meshheading:8535157-Peptide Elongation Factor Tu, pubmed-meshheading:8535157-Peptide Elongation Factors, pubmed-meshheading:8535157-Plasmids, pubmed-meshheading:8535157-Promoter Regions, Genetic, pubmed-meshheading:8535157-Protein Denaturation, pubmed-meshheading:8535157-Recombinant Proteins, pubmed-meshheading:8535157-Thermus thermophilus
pubmed:year
1995
pubmed:articleTitle
Overexpression and purification of Thermus thermophilus elongation factors G, Tu, and Ts from Escherichia coli.
pubmed:affiliation
Lehrstuhl für Biochemie, Universität Bayreuth, Germany.
pubmed:publicationType
Journal Article, Comparative Study