TABLE 1.

Strains, plasmids, and primers used in this worka

Strain, plasmid, or primerDescription or sequenceSource or reference
Strains
    MC1061F araD139 Δ(ara leu)7696 Δ(lacY74) galU galK hsdr hsdM+ strABioRad
    CL100ΔiscS17
    CL100(DE3)DE3 lysogen of CL10017
    CL201ΔcsdBThis work
    CL102Δcsd ΔcsdBThis work
    CL103ΔthiIThis work
    CL250ΔmnmAb
    CL260ΔmiaBThis work
    PK4331MG1655 lacZΔ145 ΔiscS::Knr30
    PK5930MG1655 lacZΔ145 ΔiscS::Knr ΔcsdA::Cmrc
Plasmids
    pCL010Wild-type E. coli iscS in pET21c14
    pCSDWild-type E. coli csd in pET21cThis work
    pCSDBWild-type E. coli csdB in pET21cThis work
    pKO3For deletion of genes in E. coli20
Primers
    CSD.N5′-ATC AAG CCG AGG AGT CAT ATG AAC GTT TTT AAT CCC GCG
    CSD.C5′-CGA ATT GCG GGT GAA TTC TTA ATC CAC CAA TAA TTC CAG
    CsdB.N5′-GCT GCC AGG AGG TGC CAT ATG ATT TTT TCC GTC GAC AAA
    CsdB.C5′-AGC CAT AGT GCC GGA TCC TTA TCC CAG CAA ACG GTG AAT
    CSD.No5′-AAG GAA AAA AGC GGC CGC TAC ATT TAC CCT GTC TGT CCA TAG TGAT T
    CSD.Ni5′-CAC GCA ATA ACC TTC ACA CTC CAA ATT TAT AAC CAT GGT ACT CCT CGG CTT G
    CSD.Co5′-CGC ACG CAT GTC GAC GTC TTA TCC GAC CCG GTT CT
    CSD.Ci5′-GTT ATA AAT TTG GAG TGT GAA GGT TAT TGC GTG TGA CCG CGC GCT GGA ATT A
    CsdB.No5′-AAG GAA AAA AGC GGC CGC ACG CAA CTC AAT GGC GAA AAC A
    CsdB.Ni5′-CAC GCA ATA ACC TTC ACA CTC CAA ATT TAT AAC AAT CAT CTT GCA CCT CCT GGC
    CsdB.Co5′-CGC ACG CAT GTC GAC CAT TGA CCA TCC GGC AAT GTG A
    CsdB.Ci5′-GTT ATA AAT TTG GAG TGT GAA GGT TAT TGC GTG CAC CGT TTG CTG GGA TAA CAG
  • a For 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).

  • b R. Kambampati and C. T. Lauhon, submitted.

  • c P. J. Kiley, unpublished data.