4. Evaluation of Ssp DnaE C Part Mutations Influence on the Trans-Splicing Efficiency .
| calculated
free energy changes |
experimental
data from 48-h post-transfected HEK
293 |
||||
|---|---|---|---|---|---|
| samples | affinity ΔΔG dG(kcal/mol) | stability Δstability dG(kcal/mol) | mean %GFP+ | mean MFI | mean number of GFP+ objects per image |
| pGFP | - | - | 93,7 | 236959 | 5711.78 |
| A165M | –18.92 | 0.74 | 64,0 | 11132 | 1099 |
| SSP N+C | - | - | 62,3 | 9947 | 1775 |
| A162M | –3.25 | 0.92 | 59,4 | 6250 | 992 |
| F159M | –4.61 | –2.28 | 43,4 | 1609 | 50 |
| A162F+A165M | –31.73 | –26.29 | 41,6 | 1416 | 129 |
| A162M+A165M | –22.92 | –14.04 | 26,2 | 1042 | 22 |
| I151M | –14.00 | –10.28 | 16,4 | 671 | 1.5 |
| HEK293 | - | - | 0,3 | 450 | 0.1 |
a
Experimental results after transfection of HEK293 were compared with the calculations of free energy changes in affinity and stability.