Overproduction of the Thermus thermophilus elongation factor Tu in Escherichia coli.
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.
Ahmadian
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MR
Laboratorium für Biochemie, Universität Bayreuth, Germany.
Kreutzer
R
R
Sprinzl
M
M
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*
D004269:DNA, Bacterial; D006153:Guanosine Diphosphate; D010444:Peptide Elongation Factor Tu