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Table 3 Oligonucleotide primers applied in this study

From: The predicted σ54-dependent regulator EtpR is essential for expression of genes for anaerobic p-ethylphenol and p-hydroxyacetophenone degradation in “Aromatoleum aromaticum” EbN1

Primera

Target gene

Nucleotide sequence (5' → 3')b

Product length (bp)

Gene specific primer pairs

bcrC_108_F

bcrC

CAAGTGGTGGCAACGATGTGT

191

bcrC_299_R

 

GAAGGTCTGGCGATACTGGATGC

 

pchF_1336_F

pchF

GGCCGGCAACGTCATCATC

273

pchF_1099_R

 

CCATCCGGGAGCACCACT

 

hped_153_F

hped

TGATCGAAGGCAAGGGCGGAAAAG

331

hped _473_R

 

GCGGCGGTGTAGGGCGTGATG

 

acsA_1373_F

acsA

GCCGCGGTGAGGTT

306

acsA_1068_R

 

CGGGGTGAATGTCCA

 

etpR_425_F

etpR

AATTGGCCGCTCTTCTG

310

etpR_116_R

 

TTTCGGCATGTTTGTCA

 

etpR_1717_Fc

etpR

TGGCGACGGCATTCTC

219

etpR_1499_Rc

 

TGCCGCATCTGTTCACC

 

ebA326_41_F

ebA326

TGGCTGGATCTCTGCTC

275

ebA326_315_R

 

TTCCCGTGCGACCTG

 

ebA327_1070_F

ebA327

GCTTCGCGGTCCTGA

375

ebA327_1480_R

 

TGTCGCGGTTGTAGC

 

ebA329_212_F

ebA329

TGCGGCCCCTGATG

316

ebA329_537_R

 

ACGATGCCGCTGTGG

 

ebA332_488_F

ebA332

CCGGCGTGGAGGTAG

285

ebA332_772_R

 

GGCGCGGGGTTTT

 

ebA335_1092_F

ebA335

GCTGGGGGAGACGAA

253

ebA335_1344_R

 

CGCCGCCTTGTTGT

 

Generation of Δ etpR deletion mutation

 

acsA_BamHI_F

acsA

AAGGATCCCACGAAATGTCTCCTGAACCCTGC

1300

acsA_SphI_R

 

ACCGGGCATGCGCCCACCAGC

 

ebA327_BamHI_F

ebA326/ebA327

GATCAGGATCCACGTCACCG

2350

ebA326_BamHI_R

 

AAGGATCCTGACCGTCGGAGGACCGGATAGATC

 

Identification of Δ etpR genotype

 

ΔetpR _F

3'-IR-etpR f

TGGGCGTAGCGTAGT

2133d/ 318e

ΔetpR _R

5'-IR-etpR f

TGGATTGTTCTGTAT

 

Generation of in trans complementation of etpR

 

etpR_XhoI_F

etpR

AACTCGAGCCCACTCCAAGCGTCGAAACACCGGC

2436

etpR_KpnI_R

 

AAAGGTACCGCTTCGCTCCGGGAAACCAGTGTGCGC

 
  1. aR = reverse primer; F = forward primer
  2. bRecognition sites for restriction enzymes are marked in bold type
  3. cPrimer pair applied in real time RT-PCR experiment
  4. d"A. aromaticum" EbN1 (wild type)
  5. eDeletion mutant (strain EbN1-RR001; ΔetpR genotype)
  6. fIR, intergenic region