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. Author manuscript; available in PMC: 2009 Dec 1.
Published in final edited form as: Mol Microbiol. 2008 Sep 30;70(5):1136–1151. doi: 10.1111/j.1365-2958.2008.06462.x

Table 1.

Bacterial Strains and Plasmids

Strains/Plasmids Relevant Details Reference/Source
Strains
DH5αF’IQ E. coli lacIq host strain for plasmid construction Invitrogen
SM10λpir E. coli host strain for mating plasmids into P.
aeruginosa
FW102 Escherichia coli host strain for promoter-lacZ
fusions on single copy F’ episomes bearing either a
tetracycline resistance gene (Tet) or a kanamycin
resistance gene (Kan)		 Whipple, 1998
BG77 FW102 harboring an F’ Tet bearing test promoter
placUV5C-A linked to lacZ; bears a mutation in
the chromosomal copy of rpoD specifying the
F563Y substitution linked to a kanamycin
resistance gene		 Gregory et al., 2005
FW102 OL2–62 FW102 harboring an F’ Kan bearing test promoter
placOL2-62 linked to lacZ		 Deaconescu et al., 2006
PAO1 ΔalgQ P. aeruginosa strain PAO1 containing an in-frame
deletion of the algQ gene		 This work
PAO1 ΔalgQ σ70
[I392F]		 PAO1 ΔalgQ with the chromosomal rpoD gene
specifying σ70 [I392F]		 This work
Plasmids
pACΔCI Encodes no relevant protein; used as empty vector
control; confers CamR 		Dove et al., 1997
pACRsd Encodes full-length Rsd under the control of the
lacUV5 promoter; confers CamR 		This work
pLXσ70 Encodes full-length σ70 under the control of a
weak-constitutive promoter; confers AmpR 		Gregory et al., 2005
pACλCI Encodes full-length λ CI under the control of the
lacUV5 promtoer; confers CamR 		Dove et al., 1997
pACλCI-Rsd Encodes residues 1-236 of λCI fused via three
alanine residues to full-length Rsd under the
control of the lacUV5 promoter; confers CamR 		This work
pACλCI-σ70
region 2		 Encodes residues 1-236 of λ CI fused via three
alanine residues to residues 94-448 of σ70 under
the control of the lacUV5 promoter; confers CamR
 Leibman & Hochschild, 2007
pACλCI-AlgQ Encodes residues 1-236 of λ CI fused via three
alanine residues to full-length AlgQ under the
control of the lacUV5 promoter; confers CamR 		Dove & Hochschild, 2001
pACλCI-Pa σ70
region 2		 Encodes residues 1-236 of λ CI fused via three
alanine residues to residues 96-450 of P.
aeruginosa σ70 under the control of the lacUV5
promoter; confers CamR 		This work
pBRα Encodes full-length α under the control of tandem
lpp and lacUV5 promoters; confers AmpR 		Dove et al., 1997
pBRα-σ70
region 4		 Encodes residues 1-248 of αdirectly fused to
residues 528-613 of σ70 under the control of
tandem lpp and lacUV5 promoters; bears a
mutation in the σ70 moiety specifying the D581G
substitution; confers AmpR 		Nickels et al., 2002
pBRα-σ70
region 2		 Encodes residues 1-248 of α fused via three
alanine residues to residues 94-448 of σ70 under
the control of tandem lpp and lacUV5 promoters;
confers AmpR 		This work
pBRα-Rsd Encodes residues 1-248 of α fused via three
alanine residues to full-length Rsd under the
control of tandem lpp and lacUV5 promoters;
confers AmpR 		This work
pBRα-β’
coiled-coil		 Encodes residues 1-248 of α fused via three
alanine residues to residues 262-309 of the β’
subunit of RNAP under the control of tandem lpp
and lacUV5 promoters; confers AmpR 		Leibman & Hochschild, 2007
pBRα-Pa σ70
region 2		 Encodes residues 1-248 of α fused via three
alanine residues to residues 96-450 of P.
aeruginosa σ70 under the control of tandem lpp and
lacUV5 promoters; confers AmpR 		This work
pBRα-Pa σ70
region 4		 Encodes residues 1-248 of α fused via three
alanine residues to residues 532-617 of P.
aeruginosa σ70 under the control of tandem lpp and
lacUV5 promoters; confers AmpR 		Dove & Hochschild, 2001
pCL1920 Encodes the LacZα fragment under the control of
the lac promoter; bears the pSC101 origin of
replication; confers SpecR 		Lerner & Inouye, 1990
pCLRsd Encodes full-length Rsd under the control of the
lacUV5 promoter; bears the pSC101 origin of
replication; confers SpecR 		This work
pCLAlgQ Encodes full-length AlgQ under the control of the
lacUV5 promoter; bears the pSC101 origin of
replication; confers SpecR 		This work
pLHN12His6-σ70 Encodes full-length σ70 bearing an N-terminal
hexahistidine tag under the control of a T7
promoter		 Panaghie et al., 2000
pET11aRsd-His6 Encodes full-length Rsd bearing a C-terminal
hexahistidine tag under the control of a T7
promoter		 This work
pEX18Gm Allelic replacement vector that carries colE1 origin
of replication, oriT, and sacB. Confers resistance
to gentamicin		 Hoang et al., 1998
pEXG2 Allelic replacement vector that carries colE1 origin
of replication, oriT, and sacB. Confers resistance
to gentamicin		 Rietsch et al., 2005
pM Broad host-range expression vector pMMB67EH
that carries the tac promoter, and lacIq. Confers
resistance to carbenicillin		 Furste et al., (1986)
pM-AlgQHis6 Derivative of pMMB67EH that directs the
synthesis of AlgQ-His6 under the control of the tac
promoter		 This work
pMMB67Rsd-
His6 		Derivative of pM that encodes full-length Rsd
bearing a C-terminal hexahistidine tag optimized
for codon usage (CAC) in P. aeruginosa 		This work
pPSV37 Expression vector used in P. aeruginosa. Contains
lacUV5 promoter, lacIq, colE1 and Pseudomonas
origin of replication, and oriT. Confers resistance
to gentamicin		 Arne Rietsch (Case Western
Reserve University),
unpublished
pPSV37AlgQ-
His6 		Derivative of pPSV37 that encodes full-length
AlgQ bearing a C-terminal hexahistidine tag
optimized for codon usage (CAC) in P. aeruginosa 		This work
pPSV37Rsd-His6 Derivative of pPSV37 that encodes full-length Rsd
bearing a C-terminal hexahistidine tag optimized
for codon usage (CAC) in P. aeruginosa 		This work