Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionReference or source
Strains
E. coli
  DH5αFφ80dlacZΔM15 Δ(lacZYZ-argF) recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 deoR U169Gibco BRL
  SM10Mobilizing strain, RP4 tra genes integrated in chromosome, Kmr 52
  XL1-Blue MRNonmethylating (dam dcm) cloning host used for cosmid library construction and preparation
of DNA for electroporation into B. cepacia: Δ(mcrA)183Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac
Stratagene
B. cepacia
  K56-2Clinical isolate, genomovar III 11,27
  K56-I117 pvdA::Tn5-OT182 derivative of K56-2This study
  K56-T10 pvdD::Tn5-OT182 derivative of K56-2This study
  K56pvdA::tp pvdA::Tprderivative of K56-2This study
Plasmids
 pOT182pSUP102(GM)::Tn5-OT182; Cmr Gmr AprTcr 33
 pNOT19Modified pUC19 cloning vector, Apr 47
 pUCP28TBroad-host-range vector; IncP OriT; pRO1600 ori; Tpr 50
 pPDI117-29.5-kbEcoRI fragment from I117 obtained by self-cloning, Tcr This study
 pPDI117-311.5-kbNotI fragment from I117 obtained by self-cloning, Tcr This study
 pPD519pNOT19 with 6.1-kbSphI fragment from K56-2 with pvdA geneThis study
 pPD520pUCP28T with 6.5-kb SphI fragment from pPD519This study
 p34E-TpSource of Tp cassette; Tpr 13
 pRK2013ColE1 Tra (RK2)+ Kmr 16
 pEX18TcSuicide vector; sacBTcr 23
 pZ1918Apr Gmr; source of promoterlesslacZ gentamicin resistance cassette 48
 pPD520-6ZpPD520 with 4.3-kb lacZ-Gmrcassette from pZ1918 in BglII site of pvdA gene; same orientationThis study
 pPD520-14ZpPD520 with 4.3-kblacZ-Gmr cassette from pZ1918 inBglII site of pvdA gene; reverse orientationThis study
 pUC-TPpUC-GM (46) with the gentamicin resistance cassette replaced with a 1.1-kb Tpr gene
cassette; AprTpr
This study
 pScosBC1Novel broad-host-range cosmid cloning vector based on pSuperCos1 (67); Apr Tpr This study
 pSBC-4pScosBC1 from K56-2 library containing the pvdA geneThis study
 pSBC-8pScosBC1 from K56-2 library containing thepvdA geneThis study
 pSBC-13pScosBC1 from K56-2 library containing the pvdA geneThis study