Table 1.
Comparison of TGP with GFPs for FSEC-TS assay and membrane protein expression.
| Expression host | Protein | Fusion | App. Tm (°C)a | Yield (mg L−1)b | In-gel fluorescence | FSEC | ||
|---|---|---|---|---|---|---|---|---|
| Intst. (AU) | Yield (%)c | Intst. (%) | Yield (%)d | |||||
| E. coli | aaPlsY | TGP | 91.1 | 11.1 ± 0.2 | 100 ± 15.2 | 100 ± 15.2 | 100 ± 8.4 | 100 ± 8.4 |
| ecGFP | 73.5 | 4.4 ± 0.1 | 60.6 ± 14.3 | 61.2 ± 14.5 | 55.1 ± 6.7 | 60.5 ± 6.1 | ||
| msSI | TGP | 33.3 | 16.4 ± 0.3 | 100 ± 17.0 | 100 ± 17.0 | 100 ± 15.3 | 100 ± 15.3 | |
| ecGFP | 32.0 | 2.9 ± 0.1 | 56.9 ± 7.9 | 57.5 ± 8.0 | 39.2 ± 3.6 | 43.1 ± 4.0 | ||
| S. cerevisiae | hSI | TGP | 49.4 | 1.7 ± 0.1 | 100 ± 14.5 | 100 ± 14.5 | 100 ± 4.6 | 100 ± 4.6 |
| scGFP | 44.3 | 1.6 ± 0.1 | 21.3 ± 6.7 | 46.3 ± 14.6 | 14.3 ± 0.7 | 43.3 ± 2.1 | ||
| PIS | TGP | 41.3 | 1.5 ± 0.1 | 100 ± 40.3 | 100 ± 40.3 | 100 ± 29.1 | 100 ± 29.1 | |
| scGFP | 40.5 | 1.0 ± 0.0 | 21.4 ± 4.6 | 46.6 ± 10.0 | 14.1 ± 0.8 | 42.7 ± 2.4 | ||
| Sf9 | hSI | TGP | / | 12.5 ± 0.7 | 100 ± 21.5 | 100 ± 21.5 | 100 ± 1.0 | 100 ± 1.0 |
| scGFP | / | 6.0 ± 0.2 | 17.4 ± 2.5 | 37.8 ± 5.4 | 10.9 ± 0.8 | 33.0 ± 2.4 | ||
| PIS | TGP | / | 12.5 ± 0.3 | 100 ± 24.2 | 100 ± 24.2 | 100 ± 11.2 | 100 ± 11.2 | |
| scGFP | / | 7.0 ± 0.2 | 24.7 ± 3.5 | 53.6 ± 7.5 | 15.8 ± 1.0 | 47.8 ± 3.2 | ||
| Expi293 | hSI | TGP | / | 11.1 ± 0.8 | 100 ± 12.7 | 100 ± 12.7 | 100 ± 5.4 | 100 ± 5.4 |
| scGFP | / | 1.0 ± 0.1 | 3.0 ± 0.5 | 6.5 ± 1.1 | 2.6 ± 0.1 | 7.8 ± 0.4 | ||
| PIS | TGP | / | 2.3 ± 0.5 | 100 ± 28.3 | 100 ± 28.3 | 100 ± 18.3 | 100 ± 18.3 | |
| scGFP | / | 0.6 ± 0.1 | 22.0 ± 1.6 | 47.7 ± 3.4 | 14.2 ± 1.0 | 43.0 ± 3.0 | ||
aApparent Tm was calculated by sigmoidal dose-response fitting of data from single experiments.
bYield, as mean ± standard deviation, was calculated based on the fluorescence counts of cells from three independent experiments on different colonies for bacterial and yeast expression, and on cells with different passage numbers for baculovirus and mammalian expression.
cPercent yield relative to membrane protein-TGP was calculated based on integrated arbitrary intensity (intst, AU) corrected with calibrated quantity-intensity correlation. Results (mean ± standard deviation) are from three independent experiments as in b.
dPercent yield relative to membrane protein-TGP was calculated using intensity corrected with calibrated quantity-fluorescence correlation. Data (mean ± standard deviation) are from three independent experiments as in b.