superfolder green fluorescent protein sfgfp gene  (Addgene inc)

Journal: Nature Communications

Article Title: Temporal gene regulation enables controlled expression of gas vesicles and preserves bacterial viability

doi: 10.1038/s41467-025-67667-8

Figure Lengend Snippet: Schematic representation of the genetic constructs of the dual-inducer system ( a ) and two single‑reporter constructs ( b ) for fluorescent reporter proteins expression. c , d Fluorescence fold change of dual‑inducer vs. single‑reporter across inducer concentrations. c mCherry-only vs. mCherry-dual: NS at 0 µM IPTG; P = 0.0003, <0.0001 ( P = 0.00000058), and <0.0001 ( P = 0.000003) at 20, 200, and 400 µM. d sfGFP-only vs. sfGFP-dual: NS for all. e , f Fluorescence fold change of the dual-inducer vs. single-reporter without the corresponding inducer. e mCherry-dual vs. mCherry-only. P = 0.00461, 0.0165, <0.0001 ( P = 0.00002) at 0, 50, 500 ng/mL aTc. NS at 1000 ng/mL. f sfGFP-dual vs. sfGFP-only. NS at 0, 20, 200 µM IPTG; P = 0.00461 at 400 μM. Schematic representation of the dual-inducer system induced with varying concentrations of IPTG at fixed aTc concentration ( g ) or with varying concentrations of aTc at fixed IPTG concentration ( j ). Fold change in mCherry ( h ) and sfGFP ( k ). mCherry: 0 vs. 500 ng/mL aTc across IPTG concentrations: NS at 0, 20 µM; P = 0.0238, 0.0044 at 200, 400 µM. sfGFP: 0 vs. 200 µM IPTG across aTc concentrations: NS at 0, 50 ng/mL; P = 0.0089, 0.0012 at 500, 1000 ng/mL. Fold change in sfGFP ( i ) and mCherry ( l ). i sfGFP-dual cultures at 500 ng/mL aTc without IPTG vs. 0–400 µM IPTG: NS at 0 µM; P = 0.0009, 0.0007, 0.0010 at 20, 200, 400 µM. l mCherry-dual cultures at 200 µM IPTG without aTc vs. 0–1000 ng/mL aTc: NS at 0 ng/mL; P = 0.0478, 0.0013, 0.0145 at 50, 500, 1000 ng/mL. Fold change in c – f , h , i , k , l is plotted in arbitrary units (AU, y -axis), data representing mean ± s.d. ( n = 6 biologically independent samples). Statistics: c , d , h , k by two-tailed unpaired Welch t-test; e , f , i , l vs. grey controls by Brown–Forsythe and Welch one-way ANOVA with Dunnett T3 correction.

Article Snippet: The monomeric Cherry red fluorescent protein (mCherry) gene was obtained from Addgene (plasmid #29747), and the superfolder green fluorescent protein (sfGFP) gene was acquired from Addgene (plasmid #85492).

Techniques: Construct, Expressing, Fluorescence, Concentration Assay, Two Tailed Test

Journal: medRxiv

Article Title: SARS-CoV-2 detection with de novo designed synthetic riboregulators

doi: 10.1101/2020.07.28.20164004

Figure Lengend Snippet: a) Schematics of the operating principle of toehold biosensors. b) Workflow for cell-free SARS-CoV-2 detection platform. Key target sites for detection of SARS-CoV2 were identified on the viral genome. Toehold triggers are then generated using NCBI Primer BLAST. The trigger candidates that have non-optimal free energy predictions and candidates that are not compatible with species specific amplification were eliminated. Eligible candidates were then used in the design of their respective toehold switches in silico using NUPACK©7 software. c) Validation of designed toehold switches. Triggers were cloned downstream of T7-LacO promoter. Switches were cloned downstream of T7-LacO promoter and upstream of sfGFP reporter gene. Both constructs were transformed into E. coli BL21 (DE3) cells. Cells that only have the switch plasmid and the cells that have both switch and trigger plasmids were induced with IPTG. d) Cell-free system optimization. e) POC development. Schematics of the development of the portable, low-cost electronic optical reader

Article Snippet: On the other side, pZA backbone carries a T7 promoter with sfGFP, superfolder Green Fluorescent Protein gene from the pJT119b plasmid [ ] (deposited to the Addgene by Jeffrey Tabor, Addgene #50551) and rrnB_T1 terminator from Registry of Standard Biological Parts at the downstream of sfGFP.

Techniques: Generated, Amplification, In Silico, Software, Clone Assay, Construct, Transformation Assay, Plasmid Preparation

Journal: medRxiv

Article Title: SARS-CoV-2 detection with de novo designed synthetic riboregulators

doi: 10.1101/2020.07.28.20164004

Figure Lengend Snippet: Characterization of best performing two trigger sequences found in S and ORF1ab region in vivo . For each trigger sequence, four different unique switch sequences were designed and cloned in p15A vector with T7 promoter and sfGFP coding sequence. Trigger sequences were cloned in ColE1 origin vectors with T7 promoter. Both vectors were transformed in BL21 (DE3) cells and induced. Flow cytometry results were taken after 90 minutes. Meanwhile, cells were monitored for 16 hours via total cell fluorescence measurements using a microplate reader. Cytometer results and total cell fluorescence measurements of designed switch for trigger sequence found in a) ORF1ab and b) S. For all microplate measurements, three replicates were used.

Article Snippet: On the other side, pZA backbone carries a T7 promoter with sfGFP, superfolder Green Fluorescent Protein gene from the pJT119b plasmid [ ] (deposited to the Addgene by Jeffrey Tabor, Addgene #50551) and rrnB_T1 terminator from Registry of Standard Biological Parts at the downstream of sfGFP.

Techniques: In Vivo, Sequencing, Clone Assay, Plasmid Preparation, Transformation Assay, Flow Cytometry, Fluorescence, Cytometry