Table 5.
Rates of DNA transfer to mammalian cells through T4ASS and T4BSS.
| Donor bacteria (genotype) | T4SS | Transfer system | Relaxase | Infected cells | GFP+ mammalian cells(1) |
|
|---|---|---|---|---|---|---|
| Flow cyt % | Scope | |||||
| Bh RSE247 (wt) | Functional | RSF1010 | MobA | EA.hy926 | 5.72 ± 1.37 | nq(2) |
| Bh RSE247 (wt) | Functional | RSF1010 | – | EA.hy926 | 0.29 ± 0.07 | nq(2) |
| Bh RSE247 (wt) | Functional | R388 | TrwC | EA.hy926 | 1.00 ± 0.09 | nq(2) |
| Bh RSE247 (wt) | Functional | R388 | – | EA.hy926 | 0.14 ± 0.19 | nq(2) |
| Bh RSE247 (wt) | Functional | RSF1010 | MobA | HeLa | 2.00 ± 1.48 | nq(2) |
| Bh RSE247 (wt) | Functional | RSF1010 | – | HeLa | 0.07 ± 0.05 | nq(2) |
| Bh RSE247 (wt) | Functional | R388 | TrwC | HeLa | 0.20 ± 0.03 | nq(2) |
| Bh RSE247 (wt) | Functional | R388 | – | HeLa | 0.04 ± 0.06 | nq(2) |
| Lp Lp02 (wt) | Functional | RSF1010 | MobA | CHO FcγRII | 0.35 ± 0.12 | nq(2) |
| Lp Lp03 (dotA) | No transport | RSF1010 | MobA | CHO FcγRII | 0.03 ± 0.05 | <5 × 10-6 |
| Lp Lp02 (wt) | Functional | RSF1010 | – | CHO FcγRII | 0.00 ± 0.00 | <5 × 10-6 |
| Lp CR503 (icmS icmW) | No chaperone | RSF1010 | MobA | CHO FcγRII | 0.00 ± 0.00 | <5 × 10-6 |
| Lp Lp02 (wt) | Functional | R388 | TrwC | CHO FcγRII | 0.00 ± 0.00 | <5 × 10-6 |
| Lp Lp02 (wt) | Functional | R388 | TrwC-RalF | CHO FcγRII | 0.00 ± 0.00 | 1 × 10-5 |
| Lp Lp03 (dotA) | No transport | R388 | TrwC-RalF | CHO FcγRII | nq(2) | <5 × 10-6 |
| Lp Lp02 (wt) | Functional | R388 | – | CHO FcγRII | nq(2) | <5 × 10-6 |
| Lp CR503 (icmS icmW) | No chaperone | R388 | TrwC-RalF | CHO FcγRII | nq(2) | 2 × 10-5 |
| Cb intergenic::Tn (wt) | Functional | RSF1010 | MobA | HeLa | 0.56 ± 0.53 | nq(2) |
| Cb dotA::Tn | No transport | RSF1010 | MobA | HeLa | 0.04 ± 0.02 | nq(2) |
| Cb intergenic::Tn (wt) | Functional | RSF1010 | – | HeLa | 0.10(3) ± 0.04 | <5 × 10-6 |
(1)DNA transfer is measured as the ratio of mammalian recipient cells expressing GFP. Data from flow cytometry (left column) show the percentage of GFP positive cells (mean ± SD of two to eight independent assays). Infected cells were also screened visually under the microscope (right column). Positive cells were counted and divided by the total number of cells per well (estimated as 200.000). The screen was performed at least twice for each condition. (2)nq, not quantified. (3)Due to higher background (see text for details).