Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidGenotype or descriptionPhenotypeaSource or reference
Strain
S. pneumoniae
  Cp1015Rx derivate; str1 hexA Smr 19
  Cp6602 comE38KE micB::km Kanr This work
  Cp7002 micB::km Kanr This work
  Cp7003 micB100LR This work
  Cp7009 micA59DA This work
  Cp7039 micA59DA comE::km Kanr This work
  Cp7089 micA59DA comA::ery Eryr This work
E. coli
  DH5α supE44ΔlacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 Bethesda Research Laboratories
  TOPI0F mcrAΔ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL endA nupG Invitrogen
Plasmid
 pAM239Derived from pBR322; ColE1 origin; lacZα selectionSptr 10
 pFS160.68-kb HindIII amplicon from Cp1015 containing the 16S gene inserted into pAM239Sptr 7
 pPFMD521.92-kb KpnI/HindIII amplicon (primers FMD52 and Rmic) from pPT12 inserted into pWSK29Ampr This work
 pPFMD591.90-kbKpnI/HindIII amplicon (primers FMD59 and Rmic) from pPT12 inserted into pWSK29Ampr This work
 pPFMD1051.0-kbBamHI/HindIII amplicon (primers FMD105 and Rmic) from pPT12 inserted into pWSK29Ampr This work
 pPFMG1281.0-kbBamHI/HindIII amplicon (primers FMG128 and Rmic) from pPT12 inserted into pWSK29Ampr This work
 pPFML1001.0-kbBamHI/HindIII amplicon (primers FML100 and Rmic) from pPT12 inserted into pWSK29Ampr This work
 pPFMN1201.0-kbBamHI/HindIII amplicon (primers FMN120 and Rmic) from pPT12 inserted into pWSK29Ampr This work
 pPJ1pUC derivative containing an HincII fragment carrying the kanamycin resistance geneAmprKanr 21
 pPMAE0.7-kbBamHI/EcoRI amplicon (primers FMAE and RMAE) from pPT12 containing the micA gene inserted into pTrcHis2AAmpr This work
 pPMAE590.7-kbBamHI/EcoRI amplicon (primers FMAE and RMAE) from Cp7009 containing the micA59DA gene inserted into pTrcHis2AAmpr This work
 pPMAI0.4-kbEcoRI/HindIII amplicon (primers FMAI and RMAI) from pPT12 containing an internal fragment of micAinserted into pAM239Sptr This work
 pPMBE11.23-kb BamHI/EcoRI amplicon (primers FMBE and RMBE) from pPT12 containing the micBgene inserted into pTrcHis2AAmpr This work
 pPMBE1001.23-kb BamHI/EcoRI amplicon (primers FMBE and RMBE) from Cp7003 containing themicB100LR gene inserted into pTrcHis2AAmpr This work
 pPMBEN0.88-kbBamHI/SalI DNA fragment from pPMBE1 containing the N terminal of micB inserted into pTrcHis2AAmpr This work
 pPMCI0.5-kbEcoRI/HindIII amplicon (primers ForfCin and RorfCin) from pPT14 containing an internal fragment oforfC inserted into pAM239Sptr This work
 pPT122.1-kb KpnI/HindIII amplicon (primers Fmic and Rmic) from Cp1015 containing the micAB genes inserted into pWSK29Ampr This work
 pPT12-KmpPT12micB::km AmprKanr This work
 pPT140.98-kbXbaI/XhoI amplicon (primers ForfC and RorfC) from Cp1015 containing the orfC gene inserted into pWSK29Ampr This work
 pPT14-KmpPT14orfC::km AmprKanr This work
 pPT182.9-kbXbaI-PstI fragment from Cp1015 amplified by PCR containing the comCDE operon, inserted into pWSK29Ampr 7
 pPT18-KClpPT18comE::km AmprKanr This work
 pTrcHis2ApUC-derived vector for the production and purification of recombinant proteins in E. coli Ampr Invitrogen
 pWSK29Cloning vector derived from pBlueScriptAmpr 29
 pXF208Plasmid containing an internal fragment ofcomA Emr 6
  • a Amp, ampicillin; Ery, erythromycin; Kan, kanamycin; Spt, spectinomycin.