Table 1.

Pneumococcal strains and DNA constructs

Strain, plasmid or ampliconRelevant characteristicsDescriptionSourcea or reference
Strains
 CP1250 hex malM511 str-1 bgl-1 Rx derivative, low β-galactosidase background 39
 CP1500 hex nov-r1 bry-r str-1 ery-r1 ery-r2 Donor of point markers 6
 CPM1CP1250rpoC::C-his-10::pEVP310× His tagged at C terminus of β′ of RNA polymerase with pEVP3; Cmr CP1250 × pMSL1
 CPM2CP1250 ΔcomX1::PcEmInsertion-deletion replacement of comX1 with synthetic amplicon, aMSL2, Emr CP1250 × aMSL2
 CPM3CP1250comX1′::(pEVP3)::comX1 + lacZfusion to comX1 by using pEVP3, CmrComX1+ CP1250 × pMSL2
 CPM4CPM2comX2′::(pEVP3)::′comX2 lacZfusion to comX2, ΔcomX1, ΔcomX2, Cmr Emr CPM2 × pMSL3
 CPM5CP1250 ΔcomX2::PcTetInsertion-deletion ofcomX2 with synthetic amplicon, aMSL5, Tetr CP1250 × aMSL5
 CPM6CP1250comC′::(pEVP3)::comC + lacZfusion to comC by using pEVP3, CmrComC+ CP1250 × pXF520
 CPM7CP1250ssb2′::(pEVP3)::ssb2 + lacZfusion to ssb2 by using pEVP3, CmrSsb2+ CP1250 × pMSL4
 CPM8CPM2 ΔcomX2::PcTetΔcomX1, ΔcomX2, Emr Tetr CPM2 × CPM5
 CPM9CP1250comX2′::(pEVP3)::′comX2 lacZfusion to comX2 by using pEVP3, CmrComX2+ CP1250 × pMSL3
 CPM10CPM8comC′::(pEVP3)::comC + lacZfusion to comC, ΔcomX1, ΔcomX2, ComC+ Emr TetrCmr CPM2 × CPM5 × CPM6
 CPM11CPM8comA′::(pEVP3)::′comA lacZfusion to comA, comA, ΔcomX1, ΔcomX2; Emr TetrCmr CPM2 × CPM5 × CP1649
 CPM12CPM8ssb2′::(pEVP3)::ssb2 + lacZfusion to ssb2, ΔcomX1, ΔcomX2, Ssb2+ Emr TetrCmr CPM2 × CPM5 × CPM7
 CPM13CPM8cglA′::(pEVP3)::′cglA lacZfusion to cglA, cglA, ΔcomX1, ΔcomX2; Emr TetrCmr CPM2 × CPM5 × CP1548
 CPM14CPM8celB′::(pEVP3)::′celB lacZfusion to celB, celB, ΔcomX1, ΔcomX2; Emr TetrCmr CPM2 × CPM5 × CP1601
 CPM15CPM8cflA′::(pEVP3)::′cflA lacZfusion to cflA, cflA, ΔcomX1, ΔcomX2; Emr TetrCmr CPM2 × CPM5 × CP1506
 CPM16CP1250comX2′::(pEVP3)::comX2 + lacZfusion to comX2 by using pEVP3, CmrComX2+ CP1250 × pMSL5
 CPM17CPM3 ΔcomE::PcEm lacZ fusion tocomX1, ΔcomE, EmrCmr CPM3 × aMSL6
Plasmids
 pEVP3Nonreplicative vector (Cmr lacZ) 8
 pMSL1pEVP3::′rpoC::C-His-10 Vector tagging rpoC with 10 × HispEVP3::a(DAM211, DAM254)
 pMSL2pEVP3::′ftsH comX1 comX1 targeting lacZ fusion insertionpEVP3::a(MSL13, MSL14)
 pMSL3pEVP3::′comX2Mutation ofcomX and lacZ fusionpEVP3::a(MSL27, MSL28)
 pMSL4pEVP3::ssb2 ssb2targeting and lacZ fusionpEVP3::a(DAM214, DAM215)
 pMSL5pEVP3::′nusG comX2 lacZ fusion to comX2, ComX2+ pEVP3::a(MSL42, MSL28)
 pXF520pEVP3::′orfL-comC lacZfusion to comC, ComC+ 39
Amplicons
 aMSL1PcEmEmr marker with synthetic promotera(DAM212, DAM213)
 aMSL2ComX1up::PcEm::comX1dwFor insertion-deletion replacement for comX1 See Fig. 3
 aMSL3PcConstitutive promotera(DAM212, MSL31)
 aMSL4PcTetNew synthetic markera(DAM212, MSL33)
 aMSL5ComX2up::PcTet::comX2dwFor insertion-deletion replacement for comX2 See Materials and Methods
 aMSL6ComEup::PcEm::comEdwFor insertion-deletion replacement for comE See Materials and Methods
  • a Strain construction by a cross carried out by using transformation is indicated as recipient × DNA donor × additional DNA donor.