optimized gfp gene, superfolder gfp  (GenScript corporation)

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: Analytical biochemistry

Article Title: High-throughput purification of recombinant proteins using self-cleaving intein tags.

doi: 10.1016/j.ab.2016.10.016

Figure Lengend Snippet: Fig. 6. High-throughput CBD purification results. (A) SDS-PAGE analysis of the CI-bGal purification. The purified product from 12 wells was randomly selected for analysis. The elution profile of the 116 kDa bGal protein was very similar for all 12 samples, and the purity was >90% for each. (B) SDS-PAGE analysis of the CI-sfGFP purification. The purified product from 12 wells was randomly selected for analysis. The elution profile of the 27 kDa sfGFP protein was very similar for all 12 samples, and the purity was >90% for each. (C) Individual well yields for one of the CI-bGal purification plates. Well yields range from ~13 to 19 mg. (D) Individual well yields for one of the CI-sfGFP purification plates. Well yields range from ~15 to 25 mg. (E) Heat map for one of the CI-bGal purification plates. There are no apparent patterns in the yields. Wells H1 and H7 (from the same expression well) were taken as outliers (>3s) due to no induction. (F) Heat map for one of the CI-sfGFP purification plates.

Article Snippet: For the expression of GFP fusion proteins, the gene encoding superfolder GFP (sfGFP) was amplified from the vector pET/sfGFP-LIC (Addgene plasmid 29772) using primers; F 50-AT CGT GTA CAC AAC ATG CAG GGT ATG GTG AGC AAG GGC GAG GAG-30 and R 50-GTA CAA GCT TGC CTG CAG TTA CTA CTT ATA CAG CTC GTC CAT G-3’.

Techniques: High Throughput Screening Assay, SDS Page, Expressing

Journal: Analytical biochemistry

Article Title: High-throughput purification of recombinant proteins using self-cleaving intein tags.

doi: 10.1016/j.ab.2016.10.016

Figure Lengend Snippet: Fig. 7. Fig. 6: High-throughput ELP purification results. (A) SDS-PAGE analysis of the EI-bGal purification. The purified product from 12 wells was randomly selected for analysis. The elution profile of the 116 kDa bGal protein was very similar for all 12 samples, and the purity was >90% for each. (B) SDS-PAGE analysis of the EI-sfGFP purification. The purified product from 12 wells was randomly selected for analysis. The elution profile of the 27 kDa sfGFP protein was very similar for all 12 samples, and the purity was >90% for each. The sfGFP samples run as doublets on SDS-PAGE when constituted in the ELP elution buffer, for unknown reasons. (C) Individual well yields for one of the EI-bGal purification plates. Well yields range from ~10e14.5 mg. (D) Individual well yields for one of the EI-sfGFP purification plates. Well yields range from ~7 to 14 mg. (E) Heat map for one of the EI-bGal purification plates. There are no apparent patterns in the yields. (F) Heat map for one of the EI-sfGFP purification plates. Well D12 was taken as an outlier (>3s).

Article Snippet: For the expression of GFP fusion proteins, the gene encoding superfolder GFP (sfGFP) was amplified from the vector pET/sfGFP-LIC (Addgene plasmid 29772) using primers; F 50-AT CGT GTA CAC AAC ATG CAG GGT ATG GTG AGC AAG GGC GAG GAG-30 and R 50-GTA CAA GCT TGC CTG CAG TTA CTA CTT ATA CAG CTC GTC CAT G-3’.

Techniques: High Throughput Screening Assay, SDS Page