Table 5 Bacterial strains, plasmids and primers used in this study

P. chlororaphis PA23

Phz⁺Rif^R wild type (soybean plant isolate)

[1]

PA23-443

Phz⁻ Rif^R ptrA::Tn5- OT182 genomic fusion

This study

E. coli

DH5α

supE44 ΔlacU169 (φ80 lacZ ΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1

Gibco

SM10

Mobilizing strain; RP4 tra genes integrated in chromosome; Km^R Tc^R

[33]

Plasmids

pOTI82

pSUP102(GM)::Tn5-OT182 Cm^R Gm^R Amp^R Tc^R

[34]

pOT182-443 (Xho I)

pOT182 containing ptrA::Tn5-OT182 genomic fusion

This study

pCR2.1TOPO

Cloning vector for PCR products

Invitrogen

pUCP22

Broad-host-range vector; IncP OriT, Amp^R Gm^R

[35]

pUCP22-ptrA

pUCP22 containing ptrA from P. chlororaphis PA23

This study

Primers

ptrA-F

5′-gggaaccggcttatagcca-3′

This study

ptrA-R

5′-atccagttgctggagcgtatt-3′

This study

TNP5-FORWARD

5′-accatttcaacggggtctcac-3′

[4]

TNP5-REVERSE

5′-tgactccatgtgacctccta-3′

[4]

Tn5-ON82

5′-gatcctggaaaacgggaaagg-3′

[4]

Tn5-OT182 right

5′-atgttaggaggtcacatg-3′

[4]