Figure S1. Fluorescent labeling of different bacteria with fluorescein labeled

conjugate AB2-FLgyr241. The general method is described in Materials and

Methods, as is the procedure for calculating fluoresecence intensity in different

bacteria (23). The top panel is a phase contrast image at 100x magnification

using a Nikon 80i or Nikon Eclipse Ti microscope. The middle panel is the same

image showing the fluorescence emitted from the fluorescein tagged conjugate.

The bottom panel is an overlay of the above two images, the fluorescence

emitting from the labeled conjugate is false colored as red and the background is

colored as green. The left half of the panel is the negative control and the right

half shows the bacteria treated with 1µM of AB2-FLgyr241 unless specified. Only

a few bacteria are shown in the figure. A: Staphylococcus aureus. Considerable

background fluorescence is observable. B: Salmonella. C: E. coli SM105. D: E.

coli SM105 (10µM of PPMO-FLCAT conjugate). The scale bar in the top panel is

3 µm. The procedure is described in Materials and Methods.

A B C D

Table S1. Sequence of morpholino oligonucleotides and synthetic peptides Morpholino oligogonucleotides CAT: CTGACTGAAATGCCTCACCA

Gyr241: GACCGCCGAGTCACCACCA

Gyr 313: GTTACCCTGACCGACCA

MMbla: GGTTTGAGGGACACCA

ECbla: TAAGGGCGACACACCA

SCR07 (full length) CTGTTCACTAGCTTGCAA

lac : CGGTGCGGGCCTCACCA

SCR5: CTAGC

SCR9; CACTAGCTT

SCR13: TTCACTAGCTTGC

Gyr313-14: ACCCTGACCGACCA

Gyr313-11: CTGACCGACCA

T11: TTTTTTTTTTT

Peptides AB1: YARVRRRGPRGYARVRRRGPRRC ALT14264: YARVRRRGPRR ALT14740: GPRGYARVRRRGPRR ALT14741: VRRRGPRGYARVRRRGPRR ALT14751: YARVRRRGPRGYAR AB2: YARVRRRGPRGYARVRRRGPRR AB2Fluor: Fluorescein-YARVRRRGPRGYARVRRRGPRR

Table S2. Alignment of gyrA-313 sequences. gyr-313 GTTACCCTGACCGACCA

Escherichia coli CGGTCAGGGTAAC

Acinetobacter CGGTCAGGGCAAC

Bacillus subtilis CGGTCACGGAAAC

Enterobacter cloacae TGGCCAGGGTAAC

Enterococcus faecalis CGGCCACGGAAAC

Klebsiella pneumonia CGGCAGGGTAAC

Mycobacterium marinarum CGGTCAGGGCAAC

Pseudomonas syringae CGGTCAGGGCAAC

Salmonella enterica ssp. typhimurium TGGTCAGGGTAAC

Staphylococcus aureus TGGCCAAGGTAAC

Table S3: Time course of doses with conjugates.

Assays were carried out as described in Table 1. Controls are cells with no

additional conjugate at 4 hr. Otherwise there was an additional conjugate at 4 hr.

The conjugate used was AB1gyr313. Viability of CPP alone varied between 0.5

and 0. 7.

Bacterium Conc. Assay (hr): 4 6 8 10

E. coli 5 µM 3 x 10-4 1.2 x 10-6 <10-6 6 x 10-6

No added mix “ 3 x 10-4 <10-5 9 x 10-6 6 x 10-6

B. subtilis 0.5 µM 7.3 x 10-4 2.5 x 10-5

No added mix “ 7.3 x 10-4 1.6 x 10-3

Table S4: Killing kinetics with scrambled sequence conjugates.

E. coli SM105 was the host bacterium and the assay was done after 6 h

incubation with the conjugate at 500 nM. The number following SCR is the

number of nucleotides in the conjugate. The results shown are the average of

two experiments.

Conjugate Viability

AB1 SCR 18 2.3 x 10-4

(CTGTTCACTAGCTTGCAA)

AB1 SCR 13 2.7 x 10-4

(TTCACTAGCTTGC)

AB1 SCR 9 5.6 x 10-3

(CACTAGCTT)

AB1 SCR 5 1.2 x 10-3

(CTAGC)

AB1 gyr313 1.0 x 10-3

AB2 SCR 18 6.9 x 10-4

AB2 SCR 9 2.9 x 10-4

AB2 gyr313-14 2.9 x 10-4