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Table 1 Pseudomonas strains, plasmids and oligonucleotides used.

From: The Pseudomonas putida Lon protease is involved in N-acyl homoserine lactone quorum sensing regulation

STRAINS

  

P. putida WCS358

Pseudomonas putida, wt

[33]

P. putida IBE4

lon1754::Tn5 of WCS358, Kmr

This study

P. aeruginosa 18577

PA1803lon437::ISlacz/hah of PAO1, Tcr

[34]

PLASMIDS

  

pMOSBlue

Cloning vector, Ampr

Amersham-Pharmacia

pBluescriptKS

Cloning vector, Ampr

Strategene

pRK2013

Kmr Tra+ Mob+, ColE1 replicon

[28]

pET28b

Expression vector, kmr

Novagen

pBBRmcs5

Broad-host-range vector, Gmr

[35]

pSCHIA5

pBluescriptKS carrying a 4.5 Kb BamHI/StuI fragment from IBE4 harboring some WCS358 and a fragment of Tn5, Ampr Kmr

This study

pB1A

pBluescriptKS carrying a 4.5 Kb EcoRI fragment from WCS358, Ampr

This study

pBBRlon

pBBRmcs5 carrying a 4.5 Kb EcoRI fragment from WCS358, Genr

This study

pPPUR3586H

pET28b derivative carrying ppuR of P. putida WCS358

This study

pBBRppuR

pBBRmcs5 carrying the ppuR gene amplified from pPuR3586H with primers P0FW and P0RV

This study

pPPUI220

pMP220 promoter probe vector carrying the ppuI promoter region, Tcr

[14]

pRSA220

pMP220 promoter probe vector carrying the rsaL promoter region, Tcr

[14]

OLIGONUCLEOTIDES

  

P0FW

5'-GGGTACCAATAATTTTGTTTAACTTTA-3'

[12]

P0RV

5'-GGGATCCATTGCTCAGCGGTGGCAGC-3'

[12]

FW119

5'-CATGCCATGGCCCTACTGGTAATGG-3'

This study

RV122

5'-CCCAAGCTTGGGCGTGATCGATTTTTGC-3'

This study

Tn5ext

5'-GAACGTTACCATGTTAGGAGGTC

[36]