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Table 1 Bacterial strains, plasmids and oligonucleotides used for mutagenesis.

From: Role of the ArcAB two-component system in the resistance of Escherichia colito reactive oxygen stress

Bacterial strains and plasmids

 

Characteristics

Source or reference

E. coli strains

K12

Isolate MG1655

Dr. Sydney Kustu, University of California

 

ΔarcA

ΔarcA::kan derivative of K12

This study

 

ΔarcB

ΔarcB::cm derivative of K12

This study

 

arcB::kan

derivative of K12 in which Kanr was inserted adjacent to arcB while maintaining the function of arcB

This study

 

ΔarcB-rev

kan derivative of ΔarcB with arcB::cm replaced by wild type arcB

This study

 

ΔfliC

fliC non-polar deletion mutant of K12

This study

 

ΔarcA/ΔfliC

ΔarcA::kan/ΔfliC derivative of K12

This study

Plasmids

pRB3-273C

Apr, low to medium copy number plasmid

[40]

 

pRB3-arcA

derivative of pRB3-273C containing arcA

[38]

 

pRB3-arcD2A

derivative of pRB3-arcA containing Asp54 → Ala mutation

This study

Oligonucleotides

Used for

Sequence

arcA5KO

mutagenesis of arcA

5'-tcttatcgttgaagacgagttggtaacacgcaacacgttg

aaaagtattttcgaagcggagtgtaggctggagctgcttc-3'

arcA3KO

mutagenesis of arcA

5'-tcttccagatcaccgcagaagcgataaccttcaccgtgaa

tggtggcgatgatttccggccatatgaatatcctccttag-3'

arcB5KO

mutagenesis of arcB

5'-gccctcgtcgttcttgccattgtggtacaaatggcggtaaccatggtgct

gcatggtcaggtcgaaagcattgatgttatgtgtaggctggagctgcttc-3'

arcB3KO

mutagenesis of arcB

5'-gtggcttttgccacccacgctttcagcacttctacgtcgtgacgccactc

ttctttcatctcttcaatccattcaccgaccatatgaatatcctccttag-3'

arcB-rev5

generation of arcB::kan

5'-cacattaatttttttaataaaaatggtacgcatcacacatttaactgattcatgtaacaa

atcatttaagttttgctatcttaactgcgtcatatgaatatcctccttag-3'

arcB-rev3

generation of arcB::kan

5'-gcgaatactgcgccaacaccagggaaatcttggctgcgccgtaaattattatgatga

gttacaagggcacagcactgtttttcaggccgcgtgtaggctggagctgcttc-3'

fliC5KO

mutagenesis of fliC

5'-tcgctgatcactcaaaataatatcaacaagaaccagtctgcgctgtcgag

ttctatcgagcgtctgtcttctggcttgcggtgtaggctggagctgcttc-3'

fliC3KO

mutagenesis of fliC

5'-ctgcggtacctggttagcttttgccaacacggagttaccggcctgctgga

tgatctgcgctttcgacatattggacacttcatatgaatatcctccttag-3'

  1. kan, kanamycin resistance cassette; cm, chloramphenicol resistance cassette. Sequences in bold in the table indicate those that are homologous to plasmids pKD3 and pKD4 [50], which were used as PCR templates for mutagenesis.