Table 3.
Fold induction of selected chromosomal genes of BT4001 affected by CTnDOT, as measured by quantitative RT-PCR.a
| ORF ID | Proposed function | BT4001ΩQABb | BT4007 | BT4001(pRteC)b |
|---|---|---|---|---|
| BT4762 | TraA (CTn4-bt) | 30 (± 13) | 143 (± 21) | 5 (± 2) |
| BT4763 | TraC (CTn4-bt) | 14 (± 2) | 44 (± 12) | 3 (± 2) |
| BT4764 | HP (CTn4-bt) | 96 (± 38) | 95 (± 16) | 2 (± 1) |
| BT4766 | HP (CTn4-bt) | 45 (± 13) | 163 (± 18) | 1 (± 1) |
| BT4767 | TraE (CTn4-bt) | 121 (± 23) | 257 (± 25) | 1 (± 1) |
| BT4768 | TraF (CTn4-bt) | 216 (± 10) | 350 (± 17) | 3 (± 1) |
| BT4770 | TraG (CTn4-bt) | 330 (± 13) | 531 (± 26) | 1 (± 1) |
| BT4780 | TraN (CTn4-bt) | 1 (± 9) | 116 (± 17) | 1 (± 0) |
| BT0018 | HPc | 22 (± 3) | 30 (± 10) | 2 (± 1) |
| BT0019 | HP | 24 (± 4) | 29 (± 4) | 2 (± 1) |
| BT0020 | HP | 60 (± 9) | 66 (± 21) | 2 (± 1) |
| BT0022 | HP | 9 (± 1) | 33 (± 11) | 1 (± 1) |
| BT0060 | Polysaccharide export protein | 1 (± 1) | 32 (± 7) | 1 (± 1) |
| BT0065 | α-Galactosidase precursor | 42 (± 12) | 29 (± 2) | 1 (± 1) |
| BT3135 | Integrase | 19 (± 8) | 16 (± 4) | 1 (± 1) |
| BT2118 | Transporter (AcrBDF family) | 39 (± 0) | 7 (± 0) | 1 (± 0) |
The fold induction was obtained by quantitative RT-PCR analysis. The internal standard, a constitutively expressed single-copy gene, was the σ70 gene. Triplicate experiments were analysed in three independent experiments and the results were described as fold induction (±SD). Fold induction (N) was calculated by the following formula: N = 2ΔΔCt = 2(ΔCt target-ΔCt σ70), where ΔΔCt is ΔCt target − ΔCt σ70 and ΔCt is the difference in threshold cycles for the target and the σ70 reference.
BT4001ΩQAB is BT4001 containing a single copy of the tetQ-rteA-rteB operon. BT4007 is BT4001 containing a copy of CTnDOT. BT4001(pRteC) is BT4001 containing a copy of pRteC, a plasmid containing rteC in which rteC is expressed constitutively from a heterologous promoter (Moon et al., 2005).
HP, hypothetical proteins which do not have any homologues in the databases.