Figure S1, 44.2MB tif file |
PagR purified to >95%
homogeneity. PagR was purified as described in Experimental
procedures for Electrophoretic Mobility Shift Assays and DNase 1
footprinting. Serial 1:2 dilutions of the final preparation were
electrophoresed through a 12% acrylamide SDS-PAGE gel, and percent
purity of the 44.8 pmol lane was calculated using the TotalLab TL 100
program. Valley to valley background correction was employed, and the
final percent purity of PagR was calculated to be 96.9%. MM, molecular
markers. |
Figure S2, 53.6 MB tif file |
PagR generates the same banding
pattern when assayed against two probes of the stm2344 – pagR
promoter region. Electrophoretic Mobility Shift Assays against the
probe spanning 2,457,048 nt to 2,457,454 nt of the chromosome (left box,
shown in Fig. 4) and the probe spanning from 2,457,048 nt to 2,457,339
nt of the chromosome (right box, used for DNase footprinting in Fig. 6).
0.5 pmol of fluorescent DNA probe was used for each assay. There are two
clear mobility shifts for both probes, even with 20:1 PagR:probe
ratio. |
Figure S3, 113.8 MB tif file |
Identification of PagR binding
sites in the promoter region of the stm2344-stm2340 operon.
A. Combined electropherograms from dideoxy termination Sanger
sequencing of the stm2344 promoter. Each trace is colored based
on traditional presentation of BigDye termination: adenine is green,
thymine is red, cytosine is blue, and guanine is black. B.
DNase I digestion of Pstm2344 with (black trace)
or without (orange trace) PagR present. Protected bases were identified
by aligning the sequencing traces (A) with the digestion traces
(B). |