Purification, characterization and reconstitution into membranes …
2012年4月1日 · We also constructed a plasmid pTR19-ASDSNH-cHis2R, where a DNA sequence coding for a His-tag at N-terminus of β subunit was genetically eliminated. However, the expression efficiency was more than two fold better
In this study, we have generated functional FoF1–ATP synthase whose proton-translocating c-ring is made from a single-polypeptide chain of 10 tandemly fused c subunits and discussed in terms of elastic coupling between Fo and F1. FoF1s with Fused Multimer Foc.
The Role of the βDELSEED-loop of ATP Synthase - PMC
2009年4月4日 · Bacterial Strains and Plasmids—Plasmid pTR19-ASDS, which carries the genes for the F 1 F 0-ATP synthase from thermophilic Bacillus PS3 , was used to generate mutations in the different positions of the C-terminal domain of the β subunit: Δ 380 LQDI 383, Δ 384 IAIL 387, Δ 388 GMDE 391, Δ 392 LSD 394, Δ 395-EDKL 398, Δ 399 VVHR 402, Δ ...
The β Subunit Loop That Couples Catalysis and Rotation in ATP …
2011年8月26日 · Plasmid pTR19-ASDS, which carries the genes for the F 1 F o-ATP synthase from thermophilic Bacillus PS3 (21), was used to generate deletion mutants. All deletion mutations investigated here were based on combinations of mutants from a previous study.
plasmid pTR19, the expression vector for the F 0 F 1-ATP syn-thase of thermophilic Bacillus PS3 (18), after which a His 6 tag was introduced at the N terminus of the A subunit by PCR to obtain plasmid pTR19-FABD. For the rotation assay, we used the pTR19-FABD-Avi, in which the AviTag biotinylation sequence (GLNDIFEAQKIEWHE) (19) was inserted ...
Characterization of force-induced melting using the short ssDNA ...
Characterization of force-induced melting using the short ssDNA construct (pTR19-ASDS). [A] Force-extension measurement of dsDNA (black dots) and ssDNA (green triangles). To convert dsDNA into...
Expression of TF 0 F 1 in E. coli cells. A, SDS-PAGE ... - ResearchGate
Bacterial Strains and Plasmids-Plasmid pTR19-ASDS, which carries the genes for the F 1 F o -ATP synthase from thermophilic Bacillus PS3 (31), was used to generate the mutations ␥K7stop ...
ATP Synthase with Its γ Subunit Reduced to the N-terminal Helix …
2009年9月25日 · Plasmid pTR19-ASDS, which carries the genes for the F 1 F o-ATP synthase from thermophilic Bacillus PS3 (31), was used to generate the mutations γK7stop, γQ36stop, γP43stop, and ϵI88stop.
Cooperation among c-subunits of FoF1-ATP synthase in rotation
2022年2月2日 · A plasmid for expressing the F o F 1 mutant harboring a substitution of F o-c Glu-56 with Asp (cE56D) was prepared from pTR19-ASDS (Suzuki et al., 2002) using the megaprimer method; this yielded pTR19-CE56D. The cE56D mutation …
Coupling of proton flow and rotation in the F0 motor of ATP synthase was investigated using the thermo-philic Bacillus PS3 enzyme expressed functionally in Escherichia coli cells.