Journal: The Journal of Biological Chemistry

Article Title: PhuS conformational dynamics are essential for DNA binding and heme-responsive control of the prrF operon in Pseudomonas aeruginosa

doi: 10.1016/j.jbc.2026.111314

Figure Lengend Snippet: Characterization of the heme binding properties of the PhuS R25 A variant . A , CD spectra of 10 μM apo-proteins ( left panel ) and holo-proteins ( right panel ) dialyzed in 1 mM potassium phosphate (pH 7.4). PhuS WT and PhuS R25 A spectra shown in blue and red, respectively. B , UV-Visible spectra of holo-WT and holo-R25 A protein in 20 mM Tris (pH 8.0) containing 150 mM NaCl. C , Tryptophan fluorescence quenching of WT PhuS and R25A PhuS. 1 μM of apo-WT PhuS or apo-R25 A PhuS was titrated with increasing concentrations (0 μM–12.8 μM) of heme. Samples were excited at 295 nm and emission was monitored at 337 nm. Relative binding was calculated by dividing the difference in fluorescence by the initial unbound fluorescence emission. The binding constant (K d ) was fit to a one-site binding model in Graphpad Prism by plotting relative binding as a function of heme concentration.

Article Snippet: 30 PM of oligonucleotides were added and incubated further at 37 ̊C for another 20 min. After the incubation, the protein-DNA binding reactions were run on a 10% Tris glycine polyacrylamide native gel (Bio-Rad).

Techniques: Binding Assay, Variant Assay, Circular Dichroism, Fluorescence, Concentration Assay

Journal: ACS Omega

Article Title: Instrument-Free, Low-Cost and Dual-System Detection for Norovirus Based on Self-Assembling DNA Circuitry

doi: 10.1021/acsomega.6c00618

Figure Lengend Snippet: Feasibility validation and condition optimization utilizing fluorescence. (A) Native-PAGE gel image for the CHA reaction steps. Lane 1–7: H1, H2, T, H1 + T, H2 + T, H1 + H2, H1 + H2 + T. (B) Fluorescence monitoring of the CHA reaction progress was performed at 37 °C with 60 cycles and 30 s per cycle. (C) S/N ratio and fluorescence value of experimental and control groups at various temperatures after 30 min of reaction. (D) S/N ratio and fluorescence value of experimental and control groups at various H1:H2 ratios after 30 min of reaction at 35 °C. Data with error bars are represented as means ± standard deviation (n = 3).

Article Snippet: Native PAGE gel, DNA marker, and loading buffer were ordered from Sangon Biotech Co., Ltd. (Shanghai, China).

Techniques: Biomarker Discovery, Fluorescence, Clear Native PAGE, Control, Standard Deviation