Overproduction of the Thermus thermophilus elongation factor Tu in Escherichia coli

Identification of thermodynamically relevant interactions between EF-Tu and backbone elements of tRNA

“…The current experiments examined the binding of numerous tRNA variants to each of two very similar bacterial EF-Tu sequences. Initial experiments took advantage of the availability of an E. coli strain that overexpressed the protein from T. thermophilus (Ahmadian et al, 1991), while subsequent experiments examined interactions using the T. aquaticus protein used in the co-crystal structure (Nissen et al, 1995). While these proteins are extremely similar, sharing 98 % identity, it is noteworthy that all of the known prokaryotic EF-Tu sequences share signi®cant levels of homology.…”

Identification of thermodynamically relevant interactions between EF-Tu and backbone elements of tRNA

“…The current experiments examined the binding of numerous tRNA variants to each of two very similar bacterial EF-Tu sequences. Initial experiments took advantage of the availability of an E. coli strain that overexpressed the protein from T. thermophilus (Ahmadian et al, 1991), while subsequent experiments examined interactions using the T. aquaticus protein used in the co-crystal structure (Nissen et al, 1995). While these proteins are extremely similar, sharing 98 % identity, it is noteworthy that all of the known prokaryotic EF-Tu sequences share signi®cant levels of homology.…”

Nucleotide binding and GTP hydrolysis by elongation factor Tu from Thermus thermophilus as monitored by proton NMR

“…Lowry et al (1992) have measured the 'H-I5N HMQC spectra of 15N-labeled amide groups of glycines in an overproduced . thermophilus EF-Tu-GDP (Ahmadian et al, 1991) . They observed two downfield glycine resonances at 11.9 and 11.3 ppm at 35 °C.…”

Efficient Separation of Thermus aquaticus EF-Tu Functional Complexes

“…Thermus aquaticus EF-Tu was overproduced in E. coli SCS1 carrying plasmid pTacTU2 with a Thermus aquaticus tufA gene. Homogeneous EF-Tu ‫ء‬ GDP was obtained generally according to (7,8). The purification procedure included thermal denaturation of E. coli proteins, followed by chromatographies on DEAE-Sephadex A-50 and Sephacryl S100 HR.…”

Identification of thermodynamically relevant interactions between EF-Tu and backbone elements of tRNA

“…The current experiments examined the binding of numerous tRNA variants to each of two very similar bacterial EF-Tu sequences. Initial experiments took advantage of the availability of an E. coli strain that overexpressed the protein from T. thermophilus (Ahmadian et al, 1991), while subsequent experiments examined interactions using the T. aquaticus protein used in the co-crystal structure (Nissen et al, 1995). While these proteins are extremely similar, sharing 98 % identity, it is noteworthy that all of the known prokaryotic EF-Tu sequences share signi®cant levels of homology.…”

Nucleotide binding and GTP hydrolysis by elongation factor Tu from Thermus thermophilus as monitored by proton NMR

“…Lowry et al (1992) have measured the 'H-I5N HMQC spectra of 15N-labeled amide groups of glycines in an overproduced . thermophilus EF-Tu-GDP (Ahmadian et al, 1991) . They observed two downfield glycine resonances at 11.9 and 11.3 ppm at 35 °C.…”

Efficient Separation of Thermus aquaticus EF-Tu Functional Complexes

“…Thermus aquaticus EF-Tu was overproduced in E. coli SCS1 carrying plasmid pTacTU2 with a Thermus aquaticus tufA gene. Homogeneous EF-Tu ‫ء‬ GDP was obtained generally according to (7,8). The purification procedure included thermal denaturation of E. coli proteins, followed by chromatographies on DEAE-Sephadex A-50 and Sephacryl S100 HR.…”

Identification of thermodynamically relevant interactions between EF-Tu and backbone elements of tRNA

“…The current experiments examined the binding of numerous tRNA variants to each of two very similar bacterial EF-Tu sequences. Initial experiments took advantage of the availability of an E. coli strain that overexpressed the protein from T. thermophilus (Ahmadian et al, 1991), while subsequent experiments examined interactions using the T. aquaticus protein used in the co-crystal structure (Nissen et al, 1995). While these proteins are extremely similar, sharing 98 % identity, it is noteworthy that all of the known prokaryotic EF-Tu sequences share signi®cant levels of homology.…”

Nucleotide binding and GTP hydrolysis by elongation factor Tu from Thermus thermophilus as monitored by proton NMR

“…Lowry et al (1992) have measured the 'H-I5N HMQC spectra of 15N-labeled amide groups of glycines in an overproduced . thermophilus EF-Tu-GDP (Ahmadian et al, 1991) . They observed two downfield glycine resonances at 11.9 and 11.3 ppm at 35 °C.…”

Efficient Separation of Thermus aquaticus EF-Tu Functional Complexes

“…Thermus aquaticus EF-Tu was overproduced in E. coli SCS1 carrying plasmid pTacTU2 with a Thermus aquaticus tufA gene. Homogeneous EF-Tu ‫ء‬ GDP was obtained generally according to (7,8). The purification procedure included thermal denaturation of E. coli proteins, followed by chromatographies on DEAE-Sephadex A-50 and Sephacryl S100 HR.…”