TABLE 6.
Subcellular location of different T4CP-eGFP fusion-proteins at different expression times in the absence or presence of T4SSR388.
| Expression time (h) | Without T4SS (%) |
T4SSR388 (%) |
T4SSR388 and MOBCloDF13 (%) |
|||||||
| M | 1P | 2P | M | 1P | 2P | M | 1P | 2P | ||
| TrwBR388 | 4 | 97 (77) | 11 (9) | 18 (14) | 34 (84) | 3 (8) | 3 (8) | – | ||
| 20 | 31 (15) | 155 (75) | 29 (10) | 10 (6) | 138 (90) | 6 (4) | ||||
| TMDTraJCDTrwB | 4 | 85 (83) | 11 (11) | 6 (6) | 55 (76) | 13 (18) | 4 (6) | – | ||
| 20 | 49 (40) | 62 (51) | 10 (8) | 18 (14) | 85 (67) | 23 (18) | ||||
| MobBCloDF13 | 4 | 18 (18) | 8 (8) | 73 (74) | 22 (19) | 16 (14) | 80 (68) | 7 (5) | 19 (13) | 122 (82) |
| 20 | 20 (18) | 24 (21) | 69 (61) | 16 (15) | 16 (15) | 74 (70) | 7 (4) | 43 (22) | 143 (74) | |
| MobBΔTMDa | 4 | 0 | 40 (95)b | 2 (5)b | N.d. | N.d. | ||||
| 20 | 0 | 35 (95)b | 2 (5)b | |||||||
The eGFP variants of the proteins in the first column were expressed in E. coli BL21C41 (DE3) strain by induction with 1 mM IPTG for 4 and 20 h. The numbers represent the amount of cells showing each location. Total number of cells counted for each sample was between 40 and 200. Numbers between parentheses represent the percentages calculated up to the total amount of cells counted for each sample. The main location for each condition is marked in bold. M, location through the perimeter of the cell membrane; 1P, location at a single-pole; 2P, location at both poles. –, without relevance for this study. N.d., not determined. aProtein expressed in E. coli BL21 (DE3). bPolar location in the cytosol.