Table 1.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristicsSource or reference
E. coli K-12 strains
 HB101 E. coli K-12/B hybrid; Smr recA thi pro leu hsdRM+ 5
 CC118 λpir Δ(ara-leu) araD ΔlacX74 galE galK phoA thi-1 rpsE rpoB argE(Am) recA, λpir lysogen 14
P. putida strains
 R1 (JB156)Natural isolate made Nalr 9
 SM1639 P. 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; NalrKmr This study
 SM1699 P. putida R1 (Nalr) × HB101(RK600) × CC118 λpir(pSM1621)a; P. putida R1 with mini Tn5-Km-rrnBP1-gfpmut3b*-T0-T1cassette randomly inserted into chromosome; NalrKmr This study
Plasmids
 RK600Cmr ColE1 oriV RP4oriT; helper plasmid in triparental matings 22
 pUT-mini Tn5-KmApr Kmr; transposon delivery vector for mini Tn5-Km 20
 pL1OW2NotKmr; low-copy cloning vector with pUC18-NotI polylinker 18
 pJBA25Apr; pUC18Not-RBSII-gfpmut3b*-T0-T1 J. B. Andersen
 pJBA44Apr; pUC18Not-PlacUV5-RBSII-gfpmut3b*-T0-T1 J. B. Andersen
 pSM1606Kmr; pLOW2Not-rrnBP1-RBSII-gfp(AAV)-T0-T1 This study
 pSM1621Apr Kmr; delivery plasmid for mini Tn5-Km-rrnBP1-RBSII-gfp(AAV)-T0-T1 This study
 pSM1690Kmr; pLOW2Not-rrnBP1-RBSII-gfpmut3b*-T0-T1 This study
 pSM1695Apr Kmr; delivery plasmid for mini Tn5-Km-rrnBP1-RBSII-gfpmut3b*-T0-T1 This study
  • a Triparental matings were performed as previously described (24).