Download Table S1

Table S1.
Effects of a lon mutation on antibiotic activity
Bacterial strain
tested
Halo size (diameter) against
Bacillus subtilis (mm)
Closest distance to Fusarium
oxysporum (mm)
CHA0 (wild type)
16.5 ± 1.9
7.4 ± 1.0
lon
21.1 ± 1.8*
12.9 ± 2.1*
lon+lon
14.8 ± 1.9
ND
Data represent the averages of five replicates against B. subtilis and three replicates against F.
oxysporum. ND, not determined; * indicate values that are significantly different from those in
CHA0 (wild type) at the P < 0.05 level (t-test).
Table S2.
Bacterial strains and plasmids used in this study
Strain or plasmid
Description
Source or reference
Wild type
C. Keel
DH5, HB101
Laboratory strains
W3110
Prototroph, sup°λ−
F- ompT hsdSB (rB- mB-) gal dcm (DE3)
Sambrook and Russell,
2001
Sambrook and Russell,
2001
Novagen
CHA0
Wild type
Stutz et al., 1986
CHA89
gacA::Kmr; Kmr
Laville et al., 1992
CHA1321
pycA::Sp/Sm
Takeuchi et al., 2009
CHAclpPX
clpPX
This study
CHAclpPX-lon
 clpPX-lon
This study
CHAlon
CHAlonC
lon
 lon strain with lon+ region in mini-Tn7
This study
This study
pCR-Blunt II-TOPO
Cloning vector, pUC ori; Kmr
Invitrogen
pET30b (+)
pET30-Lon
Inducible expression vector, Kmr
Novagen
pET30b (+) containing a lon region with C- This study
terminal six-His tag, Kmr
ColIbdrd-1::Tn5, IncIα, Kmr
Boulnois et al., 1985
Strains
Bacillus subtilis
M168
Escherichia coli
BL21 (DE3)
Pseudomonas protegens
Plasmids
pLG221
pME497
pME3087
pME3087clpPX
pME3087clpPX-lon
pME3087lon
Mobilizing plasmid, IncP-1, Tra, RepA(Ts); Voisard et al., 1988
Apr
Suicide vector, ColE1 replicon, Mob; Tcr
Voisard et al., 1994
pME3087 containing a BamHI/HindIII clpPX This study
region containing a deletion of 1.4 kb in the
clpPX genes; Tcr
pME3087 containing a BamHI/XbaI clpPX-lon This study
region containing a deletion of 3.9 kb in the
clpPX-lon genes; Tcr
pME3087 containing a EcoRI/XbaI lon region This study
containing a deletion of 1.2 kb in the lon gene;
Tcr
pME6031
pACYC177-pVS1 shuttle vector; Tcr
Heeb et al., 2000
pME6060
Translational aprA'-'lacZ fusion; Tcr
Blumer et al., 1999
pME6091
Transcriptional rsmZ-lacZ fusion; Tcr
Heeb et al., 2002
pME6182
pME6182lon
Mini-Tn7 gene delivery vector based on Humair et al., 2009
pME3280a,
HindIII-SmaI-KpnI-NcoI-SphI
cloning site, ColE1 replicon; Gmr Apr
pME6182 containing the lon gene in mini-Tn7 This study
pME7402
Transcriptional rsmZ-gfp fusion; Tcr
pME7699
Transcriptional rsmY-lacZ fusion in mini-Tn7 Humair et al., 2009
gene delivery vector pME6182; Gmr Apr
pUX-BF13
Helper plasmid encoding Tn7 transposon Bao et al., 1991
fusions; Apr
Dubuis et al., 2006
Table S3.
Oligonucleotides used in this study
Oligonucleotide
Description
Source or or
reference
LonUF
5'-ACGTGAATTCTGTTTACGGCCTTACGG-3',
underlining indicates the artificial EcoRI site
This study
LonUR
5'-CTTTTCCACTTGCAGCAGATCG-3',
anneals to the 5' region of LonDF
5’CGATCTGCTGCAAGTGGAAAAGTTGGCATGGACTCA
AGTGG-3'
5'-ACGTTCTAGACACTTAAGAGCCGCT-3',
underlining indicates the artificial XbaI site
This study
LonFPciI
5'-ACGTACATGTATGAAATCCCCTCGCAG-3',
underlining indicates the artificial PciI site
This study
LonRSmaI
5'-ACGTCCCGGGCTCTAAAAAGCTGTCA-3',
underlining indicates the artificial SmaI site
This study
ClpPUF
5'-ACGTGGATCCTGAATCTGACCTTCCCTGAG-3',
underlining indicates the artificial BamHI site
This study
ClpPUR
5'-ACGGAACATGCTCTGCAGTCA-3',
anneals to the 5' region of ClpXDF or LonDF2
This study
ClpXDF
5’TGACTGCAGAGCATGTTCCGTAAGCATCCGCAGCAG
GAATTC-3'
5'-ACGTAAGCTTGAAACCAAGATGGG-3',
underlining indicates the artificial HindIII site
5’TGACTGCAGAGCATGTTCCGTTTGGCATGGACTCAA
GTGG -3'
This study
LonDF
LonDR
ClpXDR
LonDF2
This study
This study
This study
This study