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Table 2 Bacterial strains, plasmids and oligonucleotide primers used in this work

From: Genetic characterization of psp encoding the DING protein in Pseudomonas fluorescens SBW25

Strain, plasmid or primer

Relevant properties

Source/reference

P. fluorescens

  

SBW25

Wild type strain isolated from sugar beet

[10]

SBW25-Sm

Spontaneous Smr derivative of SBW25

[20]

SBW25ΔdapB

DAP/lysine auxotroph of SBW25

[20]

PIL082

The hxcR-'dapB'lacZ IVET fusion strain of SBW25ΔdapB, Tcr

This work

PIL082-1

ΔdapB DUP(hxcS-psp)::pIVETD, the psp-'dapB'lacZ IVET fusion strain, Tcr

This work

SBW25-lacZ

SBW25 marked with 'lacZ in a phage locus

[28]

PBR826

Δpsp or Δpflu2427, SBW25 with a nonpolar deletion of psp, Tcs

This work

PBR827

ΔpstC or Δpflu3317, SBW25 with a nonpolar deletion of pstC, Tcs

This work

PBR828

ΔhxcR or Δpflu2424, SBW25 with a nonpolar deletion of hxcR, Tcs

This work

PBR829

DUP(hxcS-psp)::pUIC3, the psp-'lacZ fusion strain of SBW25, Tcr

This work

PBR830

DUP(pstS-pstC)::pUIC3, the pstC-'lacZ fusion strain of SBW25, Tcr

This work

Plasmid

  

pRK2013

Helper plasmid, Tra+, Kmr

[34]

pUIC3

Integration vector with promoterless 'lacZ, ori R6K, Mob+, Tcr

[21]

pIVETD

DAP-based IVET vector, pUIC3 with promoterless 'dapB'lacZ, ori R6K, Tcr

[20]

pIVETD-13

pIVETD containing 0.8 kb psp fragment fused to 'dapB'lacZ, ori R6K, Tcr

This work

pUIC3-61

pUIC3 carrying psp-'lacZ fusion, ori R6K, Tcr

This work

pUIC3-62

pUIC3 carrying pstC-'lacZ fusion, ori R6K, Tcr

This work

pUIC3-41

pUIC3 containing 1.6 kb psp deletion fragment, ori R6K, Tcr

This work

pUIC3-63

pUIC3 containing 1.6 kb pstC deletion fragment, ori R6K, Tcr

This work

pUIC3-64

pUIC3 containing 1.9 kb hxcR deletion fragment, ori R6K, Tcr

This work

Primera

  

psp-1

GAAGATCT GTTGCGTGCCCTGGAAG

 

psp-2

cagcatgcggatcc gttgacggaAGCGAGGGTCATGGATACCG

 

psp-3

tccgtcaacggatcc gcatgctgCGGCAACACCAACGTCTGC

 

psp-4

GAAGATCT GCTGCTGGAAATCCACGGT

 

hxcR-1

GAGATCT GCGGTAATGGCCACCTATG

 

hxcR-2

cagcatgcggatcc gttgacggaCGCTGCACCTCGCTGATCGA

 

hxcR-3

tccgtcaacggatcc gcatgctgTCGACGACGATGTGCGCAGC

 

hxcR-4

GAGATCT ATGCCAATTCAAACGCGCCA

 

pstC-1

GAGATCT TCACCAAGCACCTGGCGG

 

pstC-2

cagcatgcggatcc gttgacggaAGCCGTGCTTTTCCATGCCT

 

pstC-3

tccgtcaacggatcc gcatgctgTCGCTGTTTGCACCGGCCAAC

 

pstC-4

GAGATCT GCCGGTGGGCAAGACGATTTTC

 
  1. a Restriction sites incorporated into the primers are underlined. Complementary sequence designed for the SOE-PCR is indicated by small letters.