Table 2.
PrgX concentrations and plasmid copy numbers in strains used for in vivo prgQ expression studies and in OG1Sp(pCF10)
| Straina | Plasmid | Plasmid copy no./chromosome (approx no. of molecules/cell)b | PrgX |
|
|---|---|---|---|---|
| No. of molecules/cell | Concn (μM) | |||
| OG1Sp | None | 0 | 0 | 0 |
| 100-5 | None | 0 | 715 | 3.8 |
| OG1Sp | pCF10 | 2.2 (4-5) | 696 | 3.7 |
| OG1Sp | pCI3340c | 11.2 (4.6) | 0 | 0 |
a
All strains shown here are derived from OG1Sp.
b
Assuming that the average volume of the cells used in these experiments is 0.31 μM3 and that the average cell contains 2 chromosomes. All copy number determinations were carried out with at least 3 independent cultures, and at least 3 replicate qPCRs carried out on each DNA sample; the standard deviations were all <15% for pCI330 and <20% for pCF10.
c
Plasmid pBK2 and all plasmids used for in vivo analysis of PrgX repression (Fig. 4) used pCI3340 as the cloning vector; we detected no effects of cloned pCF10/prgQ DNA on copy number.