Table 1.

Strains, plasmids, and oligonucleotides

Strain, plasmid, or oligonucleotide	Descriptiona	Source or reference
E. coli
S17-1	recA pro hsdR RP4-2-Tc::Mu-kan::Tn7	ATCC
F. tularensis subsp. holarctica
LVS	Live vaccine strain	CDC
LVS-GFP	LVS carrying pFNLTP6 gro-gfp	11
LVS ΔwbtDEF	O-antigen-deficient mutant with deleted or defective wbtD, wbtE, and wbtF	This study
LVS ΔwbtDEF-GFP	LVS ΔwbtDEF carrying pFNLTP6 gro-gfp	This study
LVS WbtIG191V	O-antigen-deficient mutant with an amino acid substitution in WbtI	19
Plasmids
pSMP22-RT1	Suicide plasmid used for allelic replacement of wbtD, wbtE, and wbtF on O-antigen gene cluster; Ampr Camr	36
pFNLTP6 gro-gfp	E. coli-Francisella shuttle vector with gfp driven by Francisella groE promoter; Ampr Kanr	21
pTZ819	Complementation shuttle vector for wbtI; Ampr Kanr	19
Primers
F1	5′-ATGTAACAGGCTTAGGAAGTG-3′
F2	5′-ACGCTTGAGTTGCGCCTCCTG-3′
F3	5′-GTAAGAGGTTCCAACTTTCAC-3′
F4	5′-GAGTCTGGCTTGAGGTCTG-3′
F5	5′-GGCTTACGATAATGTTAAATTTCCTCATG-3′
F6	5′-AACAGTACTGCGATGAGTGG-3′
R1	5′-ATGATAAGCCAGAGAGACCATC-3′
R2	5′-CATTATGGTGAAAGTTGGAACC-3′
R3	5′-GCCACTCATCGCAGTACTG-3′
R4	5′-GTGACTAACAGCAATAATGATC-3′
R5	5′-ACCACTCAACAGCATGCTTTATGC-3′
R6	5′-GGCTTCACCTTCAACCCAAC-3′
Probes
Biotin-wbtE	5′-TTGACGCCGATTATTAAGGCAAGTGAGACGGTTGG-3′
Biotin-camR	5′-ATTATACGCAAGGCGACAAGGTGCTGATGCCGCTG-3′

^aAmp^r, Cam^r, and Kan^r indicate resistance to ampicillin, chloramphenicol, and kanamycin respectively.