title: Overproduction of the Thermus thermophilus elongation factor Tu in Escherichia coli.
abstract: The elongation factor Tu (EF-Tu) encoded by the tufl gene of the extreme thermophilic bacterium Thermus thermophilus HB8 was expressed under control of the tac promoter from the recombinant plasmid pEFTu-10 in Escherichia coli. Thermophilic EF-Tu-GDP, which amounts to as much as 35% of the cellular protein content, was separated from the E coli EF-Tu-GDP by thermal denaturation at 60 degrees C. The overproduced E coli-born T thermophilus EF-Tu was characterized by: i) recognition through T thermophilus anti-EF-Tu antibodies; ii) analysis of the peptides obtained by cyanogen bromide cleavage; iii) thermostability; iv) guanine nucleotide binding activity in the absence and the presence of elongation factor Ts; and v) ternary complex formation with phenylalanyl-tRNAPhe and GTP.
authors.affiliation: Laboratorium für Biochemie, Universität Bayreuth, Germany.
authors.forename: M R
authors.initials: MR
authors.lastname: Ahmadian
authors.forename: R
authors.initials: R
authors.lastname: Kreutzer
authors.forename: M
authors.initials: M
authors.lastname: Sprinzl
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D006153:Guanosine Diphosphate / Q000378:metabolism; D008969:Molecular Sequence Data; D010444:Peptide Elongation Factor Tu / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D016963:Thermus thermophilus / Q000235:genetics / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D006153:Guanosine Diphosphate; D010444:Peptide Elongation Factor Tu