Table 1

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant characteristicsaSource or reference
Strains
    P. mirabilis
     BB2000Wild type; spontaneous Rfr from PRM133
     YL1003fliL::kan-nt30; BB2000 with group II intron insertion in the nt 30 of fliL, KmrThis study
     YL1005PMI3101::kan-nt166, BB2000 with group II intron insertion in the nt 166 of PMI3101; KmrThis study
    E. coli
     XL1-BluerecA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tcr)]Stratagene
Plasmids
    pACD4K-CIntron expression vector; p15A ori, CmrSigma-Aldrich
    pYL25pACD4K-C containing retargeting intron specific to nt 30 of P. mirabilis fliL gene; CmrThis study
    pYL31pACD4K-C containing retargeting intron specific to nt 166 of PMI3101; CmrThis study
    pAR1219T7 RNA polymerase expression vector; pBR322 ori, Apr34
    pTrc99aPtrc expression vector; pBR322 ori, Apr35
    pBAD30PBAD expression vector; p15A ori, Apr36
    pBAD33PBAD expression vector; p15A ori, Cmr36
    pACYC184Cloning vector; p15A ori, Cmr, Tcr37
    pYL30pACYC184 containing a 699-bp DNA fragment containing PfliL (200-bp upstream to the start codon), the codon region and 16 bp downstream fliL; CmrThis study
    pYL35pTrc99a containing fliL coding region, AprThis study
    pYL36pBAD30 containing the Shine-Dalgarno sequence and the codon region of fliL from pYL30, AprThis study
    pYL37pBAD33 containing the Shine-Dalgarno sequence and the codon region of fliL from pYL30, CmrThis study
    pYL47pACYC184 containing PfliL (487 bp upstream to the start codon of fliL) and codon region of the complete fliLMNOPQR operon, CmrThis study
    pYL60pYL47 with deletion of HindIII fragment containing the fliQR codon region and 14 bp in the 3′ end of the flip codon region, CmrThis study
    pYL61pYL47 with deletion of the fliPQR codon region generated by PCR using pACYC184-HindIII_r and fliO-HindIII_f primers, CmrThis study
    pYL62pYL47 with deletion of the fliOPQR codon region generated by PCR using pACYC184-HindIII_r and fliN-HindIII_r primers; CmrThis study
    pYL63pYL47 with deletion of the fliNOPQR codon region generated by PCR using pACYC184-HindIII_r and fliM-HindIII_r primers, CmrThis study
    pYL64pYL47 with deletion of the fliPQR codon region generated by PCR using pACYC184-HindIII_r and fliL-HindIII_r primers, CmrThis study
    pYL65pTrc99a containing the codon region of fliLM, AprThis study
    pYL66pBAD30 containing the Shine-Dalgarno sequence and the codon region of fliLM; AprThis study
    pYL67pBAD33 containing the Shine-Dalgarno sequence and the codon region of fliLM, CmrThis study
    pGP704Suicide vector; R6K ori, Apr38
    pYL42pGP704 containing 1.5kb of PMI3099'-3101 immediately followed by fliLp::fliL from pYL30This study
    pYL70pACYC184 containing PflaA::lacZ fusion, CmrThis study
  • a Apr, Cmr, Kmr, Rfr, and Tcr indicate ampicillin, chloramphenicol, kanamycin, rifampin, and tetracycline resistance, respectively.