TABLE 1.

Strains, plasmids, and primers used in this work^a

Strain, plasmid, or primer	Description or sequence	Source or reference
Strains
MC1061	F−araD139 Δ(ara leu)7696 Δ(lacY74) galU galK hsdr hsdM+strA	BioRad
CL100	ΔiscS	17
CL100(DE3)	DE3 lysogen of CL100	17
CL201	ΔcsdB	This work
CL102	Δcsd ΔcsdB	This work
CL103	ΔthiI	This work
CL250	ΔmnmA	—b
CL260	ΔmiaB	This work
PK4331	MG1655 lacZΔ145 ΔiscS::Knr	30
PK5930	MG1655 lacZΔ145 ΔiscS::Knr ΔcsdA::Cmr	—c
Plasmids
pCL010	Wild-type E. coli iscS in pET21c	14
pCSD	Wild-type E. coli csd in pET21c	This work
pCSDB	Wild-type E. coli csdB in pET21c	This work
pKO3	For deletion of genes in E. coli	20
Primers
CSD.N	5′-ATC AAG CCG AGG AGT CAT ATG AAC GTT TTT AAT CCC GCG
CSD.C	5′-CGA ATT GCG GGT GAA TTC TTA ATC CAC CAA TAA TTC CAG
CsdB.N	5′-GCT GCC AGG AGG TGC CAT ATG ATT TTT TCC GTC GAC AAA
CsdB.C	5′-AGC CAT AGT GCC GGA TCC TTA TCC CAG CAA ACG GTG AAT
CSD.No	5′-AAG GAA AAA AGC GGC CGC TAC ATT TAC CCT GTC TGT CCA TAG TGAT T
CSD.Ni	5′-CAC GCA ATA ACC TTC ACA CTC CAA ATT TAT AAC CAT GGT ACT CCT CGG CTT G
CSD.Co	5′-CGC ACG CAT GTC GAC GTC TTA TCC GAC CCG GTT CT
CSD.Ci	5′-GTT ATA AAT TTG GAG TGT GAA GGT TAT TGC GTG TGA CCG CGC GCT GGA ATT A
CsdB.No	5′-AAG GAA AAA AGC GGC CGC ACG CAA CTC AAT GGC GAA AAC A
CsdB.Ni	5′-CAC GCA ATA ACC TTC ACA CTC CAA ATT TAT AAC AAT CAT CTT GCA CCT CCT GGC
CsdB.Co	5′-CGC ACG CAT GTC GAC CAT TGA CCA TCC GGC AAT GTG A
CsdB.Ci	5′-GTT ATA AAT TTG GAG TGT GAA GGT TAT TGC GTG CAC CGT TTG CTG GGA TAA CAG

^aFor primers, restriction sites are underlined, and the 33-bp gene replacement tag sequence is shown in bold (see text). Primers labeled No, Co, etc., were used to perform in-frame deletion by the method of Link et al. (20).

^bR. Kambampati and C. T. Lauhon, submitted.

^cP. J. Kiley, unpublished data.