TABLE 1.

Bacterial strains and plasmids used in this study

Bacterial strain or plasmid	Descriptiona	Source
Strain
2457T	Shigella flexneri serotype 2a	81
AWY3	2457T virB::Tn5; Knr	43
JAI04	BS611 (8); 2457T mxiE2::aphA-3; Knr	This work
BS103	2457T cured of the large virulence plasmid pINV	82
Plasmid
pBluescript KS(+) II	Multicopy cloning vector; Ampr	Stratagene
pBAD18	pBAD18 expression vector, pBR322 ori; Ampr	78
pBAD18-virF	pHJW4; pBAD18 expression vector carrying virF; Ampr	43
pBAD18-mxiE	pBAD18 expression vector carrying mxiE; Ampr	This work
pBAD18-mxiE-ipgC	pBAD18 expression vector carrying mxiE and ipgC; Ampr	This work
pBAD42	pBAD42 expression vector, pSC101 ori; Spcr	78
pBAD42-virF	pBAD42 expression vector carrying virF; Spcr	This work
pAFW04a	icsP promoter transcriptionally fused to lacZ in the low-copy pACYC184; Cmr. Digest with PstI/SalI or XbaI/SalI completely removes PicsP but retains lacZ and the lambda oop terminator used to prevent transcriptional read-through	39
pMCS-lacZ	pAFW04a-derived plasmid carrying the pBluescript KS(+) II multiple cloning site in place of the entire icsP promoter region; Cmr	This work
pSFUM131	IPTG inducible mxiE(1–251) and his6::ipgC in pNEB193; Ampr	This work
pPvirB(−1946)-lacZ	pAFW04a with −1946 to +54 relative to the virB +1; Cmr	This work
pPvirB(−1437)-lacZ	pAFW04a with −1437 to +54 relative to the virB +1; Cmr	This work
pPvirB(−976)-lacZ	pAFW04a with −976 to +54 relative to the virB +1; Cmr	This work
pPvirB(−402)-lacZ	pAFW04a with −402 to +54 relative to the virB +1; Cmr	This work
pPvirB(−350)-lacZ	pAFW04a with −350 to +54 relative to the virB +1; Cmr	This work
pPvirB(−250)-lacZ	pAFW04a with −250 to +54 relative to the virB +1; Cmr	This work
pPvirB(−200)-lacZ	pAFW04a with −200 to +54 relative to the virB +1; Cmr	This work
pPvirB(−116)-lacZ	pAFW04a with −116 to +54 relative to the virB +1; Cmr	This work
pPvirB(−58)-lacZ	pAFW04a with −58 to +54 relative to the virB +1; Cmr	This work
pPospD1-lacZ	pAFW04a with −1970 to +79 relative to the ospD1 +1; Cmr	44
pPospD1(mutMxiE)-lacZ	pPospD1-lacZ with the MxiE box 5′-CAATGGTTTTTTTAGAT-3′ mutated to 5′-CAATGCAAAACCATGGT-3′; Cmr	This work
pPospF-lacZ	pAFW04a with −2000 to +49 relative to the ospF ATG; Cmr	This work
pPicsA-lacZ	pMCS-lacZ with −440 to +442 relative to the icsA +1; Cmr	This work
pMAP07	pAFW04 lacking promoter sequences upstream of lacZ; Cmr	39

^aAmp^r, ampicillin resistance; Cm^r, chloramphenicol resistance; Kn^r, kanamycin resistance; Spc^r, Spectinomycin resistance.