Table 1.

Primers used in PCR-based cloning into the E. coli expression system and in insertion duplication mutagenesis

CBP genePrimera
5′3′
lytB gactggatccTAGTGATGGTACTTGGCAAGGAAAACAG actgctgcagATCTTTGCCACCTAGCTTCTCATTG
cggaattccgAGCTCTGCTATTTTTCTTAG cgggatcccgCATCTTTCCATCTTGGTCA
lytC gactggatccTGTCGCTGCAAATGAAACTGAAGTAGC gactaagcttATACCAAACGCTGACATCTACGCG
ggaattccATTAGCAAGTATCTGTTTAC cgggatcccgAGCCTTATAGTAGTTGTCA
cbpD cgaagatcttcgAAAATTTTACCGTTTATAGCA tcccccgggggaTGTCAAGGAAACTGCTTACA
ggaattcgatcTTTCTTCAACAGGTGGAACT ggaattcgatcAGCTAGAACCGTCTTTCAG
cbpE gactggatccGAATGTTCAGGCTCAAGAAAGTTCAGG gactaagcttTTCCCCTGATGGAGCAAAGTAATACC
GTATGGGAATtcTCAGGCTCAAGAAAG CAACTGGATccGTAGACAGTAATTCAT
cbpF gactggatccATTTGCAGATGATTCTGAAGGATGG tcagctgcagCTTAACCCATTCACCATTCTAGTTTAAG
cggaattccgTCTTCGGTTTGTTAGCG cggatcccgCCAATCTGGTCTAAGAG
cbpG cgcggatccgcgTATACAGATAAGAAACAAG tcccccgggggaACATTAAATCCACTCA
TTCTTGaATTcCCAAGTTGATACTTT ATAATGGatCCAACCTACCATTTATTTT
cbpI ctgaggatccGGGGATGGCAGCTTTTAAAAATC cagtaagcttGTTTACCCATTCACCATTACC
ggaattcgatcAATCCTAACAATCAATACAAG ggaattcgatcCAGTCTGCATGACACCTAA
cbpJ tcgaggatccGGTTGTCGGCTGGCAATATATCCCGT CagtaagcttCCGAACCCATTCGCCATTATAGTTGAC
ggaattcgatcCTGGCAATATATCCCGTTTC ggaattcgatcTAGATATTGCCAACCTGTTT
  • a Lowercase letters indicate added linkers used for cloning. cbpE and cbpG primers were designed such that restriction sites were incorporated into the coding sequence by changing two of the bases (lowercase letters). Sequences used for insertion duplication mutagenesis are italicized.