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Table 1 List of oligonucleotides used for this study

From: Promoter analysis of macrophage- and tick cell-specific differentially expressed Ehrlichia chaffeensis p28-Omp genes

Primers

Sequence

Orientation

Amplicon size

(bp)

Annealing temperature

(°C)

PROMOTER ANALYSIS

Gene 14-upstream sequence primers

For cloning into pPROBE-NT

RRG 183*

5' GACTCTAGAttgctcaacccataaaataatg

Forward

596

50

RRG 184

5' AGTGAGCTCtttataaaagataataaaaatttaag

Reverse

  

For cloning into pBlue-TOPO

RRG 217

5' attgctcaaccataaaataatggga

Forward

581

48

RRG 218

5' gttaataaaccttttataaaag

Reverse

  

RRG 267

5' cagttaactttctgtaaacttc

Forward

521

48

RRG 218**

 

Reverse

  

RRG 268

5' atcataagtttacaataatgtc

Forward

461

48

RRG 218

 

Reverse

  

RRG 269

5' cgttttctgctttattagaatg

Forward

400

48

RRG 218

 

Reverse

  

RRG 270

5' gttccgtatttattaatatatg

Forward

350

48

RRG 218

 

Reverse

  

RRG 271

5' catgtactgaatttgtgatttg

Forward

286

48

RRG 218

 

Reverse

  

RRG 272

5' ggataagtactttagcaagtgg

Forward

222

48

RRG 218

 

Reverse

  

RRG 273

5' taagtagtaaagttaactatag

Forward

169

48

RRG 218

 

Reverse

  

RRG 274

5' acttttgttgtaaatttgaaag

Forward

105

48

RRG 218

 

Reverse

  

RRG 217

 

Forward

516

50

IG14-35 del R

5' (PO4)-gtctagaatataaaatttctttc

Reverse

  

IG14-10 del F

5' (PO4)-taaatttttattatcttttataaaaggtttattaac

Forward

8366

56

IG14-10 del R

5' (PO4)-atgaaagaaataaagaaaagcaagtctag

Reverse

  

IG14-35 del F

5' (PO4)-ttctttatttctttcattattc

Forward

8366

48

IG14-35 del R

 

Reverse

  

IG14-10 del F

 

Forward

8343

51

IG14-35 del R

 

Reverse

  

Gene 19-upstream sequence primers

For cloning into pPROBE-NT

RRG 185

5' GACTCTAGActtttaattttattattgccacatg

Forward

334

61

RRG 186

5' AGTGAGCTCaatagtgacaaataaattaacaatag

Reverse

  

For cloning into pBlue-TOPO

RRG 185

 

Forward

308

60

RRG 445

5' atataacctaatagtgacaaataaattaac

Reverse

  

RRG 275

5' gtggcaaaagaatgtagcaataag

Forward

239

50

RRG 445

 

Reverse

  

RRG 276

5' gtgctgtttttctcacctttacac

Forward

188

63

RRG 445

 

Reverse

  

RRG 277

5' ctgacgtaatatattaaattttcc

Forward

125

55

RRG 445

 

Reverse

  

RRG 185

 

Forward

267

50

IG19-35 del R

5' (PO4)-gtcagaatataaatttttgtataaaatatcg

Reverse

  

IG19-10 del F

5' (PO4)-taatttatttgtcactattaggttat

Forward

8112

56

IG19-10 del R

5' (PO4)-gtagaagtgtcatataaaagcaag

Reverse

  

IG19-35 del F

5' (PO4)-ttatatgacacttctactattgttaatttatttg

Forward

8112

61.5

IG19-35 del R

 

Reverse

  

IG19-10 del F

 

Forward

8088

58

IG19-35 del R

 

Reverse

  

PRIMER EXTENSION ANALYSIS

Gene 14

RRG 14-5'rev

5' gccttctctgctgtcgttgattcc

 

NA

52

Gene 19

RRG 20-PEXT

5' cgttaataccactacctgctgggtcg

 

NA

58

RRG 44

5' cgcttccgtcccaattttgcttc

 

NA

58

IN VITRO TRANSCRIPTION ASSAY

Gene 14 upstream full-length+lac Z segment

RRG 217

5' attgctcaaccataaaataatggga

Forward

882

50

RRG 226

5' cgccattcgccattag

Reverse

  

RRG 218

5' gttaataaaccttttataaaag

Forward

882

50

RRG 226

 

Reverse

  

Gene 19 upstream full-length+lac Z segment

RRG 217

5' attgctcaaccataaaataatggga

Forward

601

50

RRG 226

 

Reverse

  

RRG 445

5' atataacctaatagtgacaaataaattaac

Forward

601

50

RRG 226

 

Reverse

  

IN VITRO TRANSCRIPTION COUPLED TRANSLATION ASSAY

RRG 185

5' gactctagacttttaattttattattgccacatg

Forward

848

58

RRG 247

5' tccggctcgtatgttgtgtg

Reverse

  
  1. * Text in capital letters refers to sequences inserted for creating restriction enzyme sites. ** Primer sequences were presented only once when a primer was described for the first time.