Table 1.

Bacterial strains and plasmids

Strain or plasmidRelevant characteristicsaSource or reference
P. putida
  G7Wild-type strain containing the NAH7 catabolic plasmid 3
  G7.C1NAH7-cured derivative of G7D. T. Gibson
  G7 Y1 nahY::Km mutantThis work
E. coli
  DH5αFλ recA1Δ(lacZYA-argF)U169 hsdR17 thi-1 gyrA96 supE44 endA1 relA1 φ80lacZΔM15Gibco-BRL
  S17-1 thi pro hdsR hdsM+ recAchromosomal insertion of RP4-2(Tc::Mu Km::Tn7) 14
 pBBR1-MCS2Kmr; multicopy broad-host-range vector 9
 pBBR1-MCS5Gmr; multicopy broad-host-range vector 9
 pLAFR1Tcr; low-copy-number broad-host-range cosmid 4
 pSUP102::GmGmr Cmr; suicide vector in Pseudomonas 13
 pUC19Apr; ColE1 replicon 19
 pHG59Kmr; pBBR1-MCS2 containing a 5.9-kbEcoRI fragment from NAH7b This work
 pHG60Kmr; pBBR1-MCS2 containing a 4-kbSmaI-EcoRI fragment from pHG59 cloned intoHincII-EcoRI sitesThis work
 pHG70Apr; the insert of pHG60 is cloned into theKpnI-BamHI sites of pUC19, resulting inEcoRI sites on either side of the insertThis work
 pHG95Apr Kmr; pHG70 with the 3′ end of nahY removed with NsiI and replaced with a GenBlock Kmr cassette from Pharmacia that had been digested with PstIThis work
 pHG97KmrGmr; the EcoRI insert of pHG95 (containingnahY::Km) cloned into pSUP102::GmThis work
 pHG100Tcr; pLAFR1 containing a 25-kb EcoRI fragment from NAH7b 5
 pHG125Gmr; 1.7-kb BsiWI fragment from pHG59 that contains nahY introduced into theAcc651 (KpnI) site of pBBR1-MCS5This work
  • a Apr, ampicillin resistant; Gmr, gentamicin resistant; Cmr, chloramphenicol resistant, Tcr, tetracycline resistant.

  • b See Fig. 1.