TABLE 1

The Chlamydia molecular genetic toolbox

Plasmid category and nameParental plasmidSelectable marker(s)Fluorophore(s)aApplication(s)FeatureReference
Plasmids for ectopic gene expression
    p2TK2-SW2 plasmid series
        p2TK2-SW2pGFP-SW2blaNoneEctopic expression from native promoterMultiple-cloning site20
        p2TK2-SW2 IncDProm-RSGFP-IncDTermp2TK2-SW2blaRSGFPFluorescent bacteria; ectopic gene expression from incD promoterGene expression from incD promoter20
        p2TK2-SW2 IncDProm-mCherry-IncDTermp2TK2-SW2blamCherryFluorescent bacteria; ectopic gene expression from incD promoterGene expression from incD promoter20
        p2TK2-SW2 IncDProm-CFP-IncDTermp2TK2-SW2blaCFPFluorescent bacteria; ectopic gene expression from incD promoterGene expression from incD promoter20
    pBOMB4 vector series
        pBOMB4pL2 [C. trachomatis LGV L2 (434/Bu)]blaGFPFluorescent bacteria; ectopic gene expression from native promotersMultiple-cloning site21
        pBOMB4-MCIpL2 [C. trachomatis LGV L2 (434/Bu)]blamCherryFluorescent bacteria; ectopic gene expression from native promotersMultiple-cloning site21
        pBOMB4RpL2 [C. trachomatis LGV L2 (434/Bu)]blaGFPFluorescent bacteria; ectopic gene expression from rpoB promoterrpoB promoter upstream of multiple-cloning site21
        pBOMB4R-MCIpL2 [C. trachomatis LGV L2 (434/Bu)]blamCherryFluorescent bacteria; ectopic gene expression from rpoB promoterrpoB promoter upstream of multiple-cloning site21
    Tetracycline-inducible vectors
        pBOMB4-Tet-mCherrypL2 [C. trachomatis LGV L2 (434/Bu)]blaGFP, mCherryFluorescent bacteria; inducible ectopic gene expressionTetracycline-inducible promoter21
        pASK-GFP/mKate2-L2pL2 [C. trachomatis LGV L2 (434/Bu)]blaRSGFP, mKate2Fluorescent bacteria; inducible ectopic gene expressionTetracycline-inducible promoter26
    Plasmids for blasticidin/chloramphenicol selection
        pGFPBSD/Z:SW2pGFP-SW2bsd, bleRSGFPFluorescent bacteria; ectopic gene expression from native promotersBlasticidin resistance selectable marker27
        pGFP-CAT::SW2pGFP-SW2catRSGFPFluorescent bacteria; ectopic gene expression from native promotersChloramphenicol resistance selectable marker28
Plasmids for targeted gene disruption
    pDFTT3-bla (TargeTron)pACD4K-Cbla (GII intron), catNoneGene disruption in C. trachomatis LGV L2incA (C. trachomatis LGV L2) gene disruption33
    pDFTT3-aadA (TargeTron)pDFTT3-blaaadA (GII intron), catNoneGene disruption in C. trachomatis LGV L2Spectinomycin resistance53
  • a RSGFP, red-shifted GFP.