Table 3.

Bacterial strains and plasmids

Strain	Relevant genotype/phenotype	Reference
E.coli
DH5α	F-endA1 hsdR17 (r- m+) supE44 thi-1 recA1 gyrA/ (Nalr) relA1 Δ(lacZYA-argF)U169Ф80dLacZΔM15λpirRK6	Laboratory collection
DY330	W3110 ΔlacU169 gal490 λcl1857 Δ(cro-bioA)	(Yu et al, 2000)
L. pneumophila
MB110	Lp02 wild-type, StrR, Thy-, HsdR-	(Berger & Isberg, 1993)
MB699	Lp02 ΔdksA::FRT::cat::FRT mutant	This work
MB697	Lp02 relA::gent spoT::kan double mutant	(Dalebroux et al, 2009)
MB698	Lp02 p206-gent, vector control strain	This work
MB701	Lp02 ΔdksA::FRT::cat::FRT mutant p206-gent	This work
MB731	Lp02 ΔdksA::FRT::cat::FRT pdksAi-gent	This work
MB698	Lp02 p206-cat, vector control strain	(Dalebroux et al, 2009)
MB686	Lp02 relA::gent spoT::kan double mutant p206-cat	(Dalebroux et al, 2009)
MB687	Lp02 relA::gent spoT::kan double mutant prelAL.p.	(Dalebroux et al, 2009)
MB700	Lp02 relA::gent spoT::kan double mutant pdksAi-cat.	This work
MB359	Lp02 prelAE.c.	(Hammer & Swanson, 1999)
MB702	Lp02 ΔdksA::FRT::cat::FRT mutant prelAE.c.	This work
MB355	Lp02 pflaAgfp td(Δ)I	(Hammer & Swanson, 1999)
MB732	Lp02 ΔdksA::FRT::cat::FRT mutant pflaAgfp td(Δ)I	This work
MB685	Lp02 relA::gent spoT::kan double mutant pflaAgfp td(Δ)I	(Dalebroux et al, 2009)
MB733	Lp02 pfliAgfp td(Δ)I	This work
MB735	Lp02 ΔdksA::FRT::cat::FRT mutant pfliAgfp td(Δ)I	This work
MB734	Lp02 relA::gent spoT::kan double mutant pfliAgfp td(Δ)I	This work
Plasmids
pGEMT-Easy	MCS within coding region of B-lactamase α-fragment linearized with single-T overhanges, AmpR	Promega
pGEM-Gent	pGEMT-Easy with 1.7 kb gentamycin cassette cloned into the MCS, source of 1.7 kb gentamycin resistance cassette, AmpR, GentR	(Molofsky et al, 2005)
pKD3	Template plasmid for λ Red system, bla FRT::cat::FRT, ori R6K	(Datsenko & Wanner, 2000)
pGEM-dksA	pGEMT-Easy with 1.4 kb PCR amplified dksA chromosomal region ligated into T overhangs, AmpR	This work
pGEM-ΔdksA::FRT::cat::FRT	pGEM-dksA with 1.1 kb FRT::cat::FRT cassette from pKD3 recombineered using strain DY330 to delete and replace the dksA coding sequence leaving ~500 bp homology on either side for Lp02 chromosomal recombination, AmpR , CamR	This work
pGEM-dksAi	pGEMT-Easy with 540 bp PCR amplified dksA chromosomal region ligated into T overhangs, AmpR	This work
p206-cat	pMMB66EH derivative, Δmob lacIq,PtaclacUV5, CamR	(Morales et al, 1991)
pdksAi-cat	p206-cat with 540 bp SalI/HindIII fragment from pGEM-dksAi ligated between the SalI/HindIII sites in the MCS, colinear with the PtaclacUV5 promoter, lacIq, Inducible dksA expression, CamR.	This work
p206-gent	p206-cat with the cat cassette replaced by the gentamycin resistance cassette from pGEM-Gent, GentR.	This work
pdksAi-gent	p206-gent with 540 bp SalI/HindIII fragment from pGEM-dksAi ligated between the SalI/HindIII sites in the MCS, colinear with the PtaclacUV5 promoter, lacIq, Inducible dksA expression, GentR.	This work
prelAE.c.	pMMB66EH derivative, lacIq,PtaclacUV5, GentR with a relA allele from E. coli encoding a truncated, metabolically active RelA, cloned into the MCS, colinear with the PtaclacUV5 promoter, lacIq Inducible relAE.c. expression, GentR.	(Hammer & Swanson, 1999)
pflaAgfp	150 bp flaA promoter fragment fused to GFPmut3 in pKB5 with Ptac and lacIq removed, td(Δ)i	(Hammer & Swanson, 1999)
pfliAgfp	304 bp fliA promoter fragment fused to GFPmut3 in pKB5 with Ptac and lacIq removed, td(Δ)i	This work