TABLE 1

Strains

Strain, cosmid, or plasmidRelevant genotype (phenotype)Sourcea
R. sphaeroides strains
    2.4.1Wild type
    2.4.1ΔS2.4.1 lacking 42-kb plasmidSuwanto and Kaplan (1992)
    CM012.4.1ΔS CI trpE::Tn5TpMCS (Tpr Trp-)This work
    CM022.4.1ΔS CI trpD::Tn5TpMCS (Tpr Trp-)This work
    CM032.4.1ΔS CI ybaU::Tn5TpMCS (Tpr Trp-)This work
    CM04b2.4.1ΔS CI argDF::Tn5TpMCS (Tpr Arg-)This work
    CM052.4.1ΔS CI ybaU::Tn5TpMCS (Tpr Trp-)This work
    CM062.4.1ΔS CII trpB::Tn5TpMCS (Tpr Trp-)This work
    CM07CM06 complemented with cosmid pUI8536 (Tpr Tcr Trp+)This work
    CM082.4.1 aroR::Ω Sm/Sp (Smr Spr Trp+)This work
    CM092.4.1 CI trpA::Ω Sm/Sp (Smr Spr Trp-)This work
    CM10CM09 complemented with cosmid pUI8063 (Tcr Smr/Spr Trp+)This work
    CM112.4.1 trpE::Ω Sm/Sp (Smr Spr Trp-)This work
    CM122.4.1 trpG::Ω Sm/Sp (Smr Spr Trp-)This work
E. coli strains
    S17-1thi pro hsdR hsdM+ recA integrated plasmid RP4-Tc::Mu-Km::Tn7Simon et al. (1983)
    DH5αF-, ϕ80ΔlacZΔM15 Δ(lacZYA-argF)U169 recA endA hsdR supE44 thi gyrA relA
    DH5α-Phe-F-, ϕ80ΔlacZΔM15 Δ(lacZYA-argF)U169 recA endA hsdR supE44 thi gyrA relA phe::Tn10dCm, CmrJ. Eraso (University of Texas Medical School)
    XL-Blue MRAΔ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 gyrA96 relA1 lacStratagene
Cosmid and plasmid vectors
    pLA2917TcrAllen and Hanson (1985)
    pLAFRxDerivative of pLAFR ori RK2 (Mob+ Tcr)Heath et al. (1992)
    pSUPTn5TpMCSTn5 derivative (Tpr, unique EcoRI site and sites for AseI, DraI, SnaBI, and SpeI)Choudhary et al. (1994)
    pBluescript SK (+) (pBS)T3 and T7 promoters (Apr)Stratagene
    pBluescript II KS (+) (pBSII)T3 and T7 promoters (Apr)Stratagene
    pUI1638Source of 2082-bp Ω Sm/Sp, see GenBank accession number M60473 (Apr Smr Spr)
    pSUP203pBR325 derivative (Mob+ Apr Cmr Tcr)
Constructs
    pUI8063pLA2917 derivative containing CI trpA region (Tcr)This work
    pUI8536pLA2917 derivative containing CII trpFB region (Tcr)This work
    pUI8668pLA2917 derivative containing CI region helO-trpEGD region (Tcr)This work
    pLX1P20pLAFRx derivative containing CI trpEGDC (Tcr)This work
    pCM01cpBSII containing a 7.1-kb Rs-Tn5 EcoRI fragment from CM01 (Tpr Apr)This work
    pCM02cpBSII containing a 17.4-kb Rs-Tn5 EcoRI fragment from CM02 (Tpr Apr)This work
    pCM02SalpBSII containing a 24-kb SalI fragment from CM02 (Tpr Apr)This work
    pCM03cpBSII containing a 10-kb Rs-Tn5 EcoRI fragment from CM03 (Tpr Apr)This work
    pCM04cpBS containing an 11-kb Rs-Tn5 EcoRI fragment from CM04 (Tpr Apr)This work
    pCM05cpBSII containing a 16.5-kb Rs-Tn5 EcoRI fragment from CM05 (Tpr Apr)This work
    pCM06cpBS containing a 7.0-kb Rs-Tn5 EcoRI fragment from CM06 (Tpr Apr)This work
    pCM07ApBS containing a 6.5-kb BamHI fragment containing trpA from cosmid pUI8063 (Apr)This work
    pCM08AInsertion of a 1.3-kb SmaI fragment from pCM07A containing trpA at EcoRV site of pBS (Apr)This work
    pCM09ApCM08A containing Ω Sm/Sp inserted into filled StyI site within trpA (Apr Smr Spr)This work
    pCM10APvuII fragment trpA::Ω Sm/Sp from pCM09A inserted into ScaI site of pSUP203 (Mob+ Tcr Smr Spr)This work
    pCM11EpBSII KS containing a 2.9-kb PstI fragment containing trpE from pCM03This work
     pCM12E1.4-kb HindIII/HincII fragment (containing trpE) from pCM11E inserted into pBSII KS (Apr)This work
    pCM13EpCM12E with Ω Sm/Sp inserted into BstEII site in trpE (Apr Smr Spr)This work
    pCM14EPvuII fragment containing trpE::Ω Sm/Sp from pCM13E inserted into ScaI site of pSUP203 (Mob+ Tcr Smr Spr)This work
     pCM15G2.3-kb PstI fragment from pCM03 inserted into PstI site of pBSIIThis work
    pCM16GpCM15G with Ω Sm/Sp inserted into NsiI site within trpG (Apr Smr Spr)This work
    pCM17GPvuII fragment trpG::Ω Sm/Sp from pCM16G inserted into ScaI site of pSUP203 (Mob+ Tcr Smr Spr)This work
    pCM18R1.3-kb BglII fragment containing aroR from CII cosmid pUI8536 inserted into BamHI site of pBS (Apr)This work
    pCM19RpCM18R with Ω Sm/Sp inserted into SmaI site within aroR (Apr Smr Spr)This work
    pCM20RpCM19R PvuII fragment aroR::Ω Sm/Sp inserted into ScaI site of pSUP203 (Mob+ Tcr Smr Spr)This work
  • a Unless otherwise stated, strains shown were generated in this study.

  • b Tn5TpMCS insertion is located between argD and argF.

  • c Rs-Tn5 indicates a R. sphaeroides-Tn5 hybrid fragment.