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Table 3 PCR primers used to create T4ΩRD mutants and the corresponding genes deleted

From: Regions of Diversity 8, 9 and 13 contribute to Streptococcus pneumoniae virulence

 

Genes deleted: TIGR annotation

Primers used to amplify flanking DNA fragments for PCR construct deletion mutagenesis†

Strain

 

Upstream fragment

Downstream fragment

T4 ΩRD2

SP0163-0168

SP0162 F: NNNNNaagcttGGGATTGGGCTCCTATG

SP0168 F: NNNNNgcggccgcGTACCAACAAATACCTC

  

SP0163 R: NNNNNactagtCTGCTAAAGATAGG

SP0170 R: NNNNNtctagaTCTAACTCTTCATAG

T4 ΩRD5

SP0694-0700

SP0694 F: NNNNNaagcttCCTATCTTGTTGTCATTA

SP0698 F: NNNNNgcggccgcATGGGAACTACTCC

  

SP0694 R: NNNNNactagtCGTAGAACATATTTCGTCC

SP0800 R:NNNNNtctagaACCCAAAACTTCTGC

T4 ΩRD7

SP1130-1145

SP1130 F: NNNNNaagcttCAATTTCTGAAGGTCCG

SP1145 F: NNNNNgcggccgcCATACTGATGATGAGG

  

SP1130 R: NNNNNactagtGTTTAAGCCATGGCG

SP1146 R: NNNNNtctagaCCATAACTAACTCTCC

T4 ΩRD8

SP1315-1352

SP1315 F: NNNNNgcggccgcACTTCGGAAAGAAGTGG

SP1352 F: NNNNNaagcttGTTTCAATGCCCAGCTTCGTCC

  

SP1315 R: NNNNNtctagaCTTCACTTTCATAATACG

SP1352 R: NNNNNactagtCTTTAATGCATCATTAACGACGC

T4 ΩRD8a

SP1315-1331

SP1315 F

SP1331 F: NNNNNaagcttTGTTACTGCAAAAAGAAC

  

SP1315 R

SP1331 R: NNNNNactagtGCGCCTTTATCTGCAGC

T4 ΩRD8b

SP1332-1352

SP1332 F: NNNNNgcggccgcGAATTTCTTTGTATCGG

SP1352 F

  

SP1332 R: NNNNNtctagaGCAGCAAACTATAAAC

SP1352 R

T4 ΩRD8b1

SP1332-1337

SP1332 F

SP1337 F: NNNNNaagcttCAGTTTACCAAATCATC

  

SP1332 R

SP1337 R: NNNNNactagtCTGCCTCTTCAGAACAATAACG

T4 ΩRD8b2

SP1338-1344

SP1339 F: NNNNNgcggccgcTGCGGTAACTGGAGTG

SP1344 F: NNNNNaagcttTAAAATATACTGGCACGG

  

SP1338 R: NNNNNtctagaTTCTTCTCTACAAGCTC

SP1344 R: CGATGTAGTTactagtGCC

T4 ΩRD8b3

SP146-1352

SP1346 F: NNNNNgcggccgcCCAACAAGTAAATGTCC

SP1352 F

  

SP1346 R:NNNNNtctagaTATAGCGCACCATACC

SP1352 R

T4 ΩRD9

SP1615-1621

SP1615 F: NNNNNaagcttGGGACTTAACTTGAGC

SP1621 F: NNNNNgcggccgcATAGGGAATGTTTACCC

  

SP1615 R: NNNNNactagtCCATTCAGGAGTTGCC

SP1621 R: NNNNNtctagaAAACAAGAAC

T4 ΩRD12

SP1950-1957

SP1950 F: NNNNNaagcttACGAGTGGATTGAC

SP1957 F: NNNNNgcggccgcGATTGAACTTAAACAGG

  

SP1950 R: NNNNNactagtCTTCATAGCTGTGATCCG

SP1957 R: NNNNNtctagaGCCATCTCCCAAATTGCC

T4 ΩRD13

SP2158-2166

SP2158 F: NNNNNaagcttGGAATTCTCAACATAG

SP2166 F: NNNNNgcggccgcCATCTCCTCATCAGG

  

SP2158 R: NNNNNactagtCCACAAATCTTTGC

SP2166 R: NNNNNtctagaCAGATTTGACGAAAGG