Tabel III.
Amount of Protein After IMAC Purification Using 25 wt and Sequence-Optimized Genes for Expression in E. coli
| Protein yield (mg/L) | Purification mode | Yield ratio after purification | Ratio total expression | ||
|---|---|---|---|---|---|
| Name | Wild-type gene | Optimized gene | Native (N)/ denat. (D) | opt/wt (elution fractions) | opt/wt (lysate) |
| Pim-1 | 10.5 | 19.4 | N | 1.85 | 1.65 ▴ |
| Pim-2 | n.d. | 10 | N | 1.29 ▴ | |
| MAPKAPK5 | 14.9 | 80 | N | 5.37 | 2.77 ▴ |
| TRIB1 | 11.3 | 6.3 | N | 0.56 | 0.83 ▾ |
| FYN | n.d. | 6 | N | 0.96 ▸ | |
| MAPK1 | 24.3 | 11.5 | N | 0.47 | 0.38 ▾ |
| MAPK14 | n.d. | 43 | N | 1.52 ▴ | |
| MAPK8 | 33.8 | 42.2 | N | 1.25 | 0.80 ▾ |
| EMG1 | 30 | 31.5 | N | 1.05 | 1.27 ▴ |
| IFN gamma | 76 | 22 | N | 0.29 | 0.53 ▾ |
| IFN alpha | 0 | 13.3 | N | 5.00 ▴ | |
| TNF alpha | n.d. | 68 | N | 1.56 ▴ | |
| NFKB1a | 34 | 34.8 | N | 1.02 | 1.11 ▴ |
| YY1 | n.d. | 24 | N | 2.57 ▴ | |
| TFIIB | 19.1 | 4 | N | 0.21 | 0.83 ▾ |
| Kif11 | 2 | 3.5 | N | 1.75 | 2.33 ▴ |
| CDC2 | 8.2 | 14 | N | 1.71 | 3.47 ▴ |
| Caveolin 1 | 0 | 1.2 | N | 10.95▴ | |
| CSF2 | 8.5 | 23.8 | D | 2.80 | 1.72 ▴ |
| IL-4 | 25.5 | 34.8 | D | 1.36 | 1.40 ▴ |
| IL-6 | 19.6 | 21.2 | D | 1.08 | 1.10 ▴ |
| IL-7 | 0 | 16.2 | D | 2.00 ▴ | |
| IL-10 | 6 | 5.8 | D | 0.97 | 1.00 ▸ |
| IFN alpha | 0 | 25.5 | D | 5.00 ▴ | |
| TNF alpha | 25.5 | 38.6 | D | 1.51 | 1.56 ▴ |
| CCL5 | n.d. | 63 | D | 2.72 ▴ | |
| TFIIB | 25.9 | 19 | D | 0.73 | 0.83 ▾ |
Columns are (left to right): Name: gene symbol; wt gene: amount of protein quantified after expression and purification using the wt coding sequence; optimized gene: amount of protein quantified after expression and purification using the optimized coding sequence; n.d.: not determined; native/denat.: purification performed under native or denaturing conditions; yield ratio (opt/wt): factor calculated from protein yield in purification elution fractions; ratio total expression (opt/wt): factor calculated from expression level in crude lysates (see Table I).