Table 1.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristicsSource or reference
E. coli K-12 strains
 HB101E. coli K-12/B hybrid; SmrrecA thi pro leu hsdRM+5
 CC118 λpirΔ(ara-leu) araD ΔlacX74 galE galK phoA thi-1 rpsE rpoB argE(Am) recA, λpir lysogen14
P. putida strains
 R1 (JB156)Natural isolate made Nalr9
 SM1639P. putida R1 (Nalr) × HB101(RK600) × CC118 λpir(pSM1695)a; P. putida R1 with mini Tn5-Km-rrnBP1-gfp(AAV)-T0-T1cassette randomly inserted into chromosome; NalrKmrThis study
 SM1699P. putida R1 (Nalr) × HB101(RK600) × CC118 λpir(pSM1621)a; P. putida R1 with mini Tn5-Km-rrnBP1-gfpmut3b*-T0-T1cassette randomly inserted into chromosome; NalrKmrThis study
Plasmids
 RK600Cmr ColE1 oriV RP4oriT; helper plasmid in triparental matings22
 pUT-mini Tn5-KmApr Kmr; transposon delivery vector for mini Tn5-Km20
 pL1OW2NotKmr; low-copy cloning vector with pUC18-NotI polylinker18
 pJBA25Apr; pUC18Not-RBSII-gfpmut3b*-T0-T1J. B. Andersen
 pJBA44Apr; pUC18Not-PlacUV5-RBSII-gfpmut3b*-T0-T1J. B. Andersen
 pSM1606Kmr; pLOW2Not-rrnBP1-RBSII-gfp(AAV)-T0-T1This study
 pSM1621Apr Kmr; delivery plasmid for mini Tn5-Km-rrnBP1-RBSII-gfp(AAV)-T0-T1This study
 pSM1690Kmr; pLOW2Not-rrnBP1-RBSII-gfpmut3b*-T0-T1This study
 pSM1695Apr Kmr; delivery plasmid for mini Tn5-Km-rrnBP1-RBSII-gfpmut3b*-T0-T1This study
  • a Triparental matings were performed as previously described (24).