Table 2

PCR primers used in this study (from Invitrogen)

Primer nameGeneration ofSequence (5′–3′)
catforwpIJ790 ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTtcgggcacgtaagaggttcc
catrevpIJ790 TTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGAttaagggcaccaataactgc
apraforwpIJ773 tcatgagctcagccaatc
oriTrevpIJ773 cgccagcctcgcagagcag
Sc9B1.20forwcyc2 deletion CGGCGGATGCGGTCGAAGAGCCCTGGGTAGGGCCGGGCCattccggggatccgtcgacc
Sc9B1.20revcyc2 deletion CGAGCCACGAAAGAGTGAGACTGAACGTCCGTCAGCGCGtgtaggctggagctgcttc
Sc9B1.20ΔNforwcyc2ΔN deletion AGCCCTGGGTAGGGCCGGGCCATGACGCAACAGCCCTTCattccggggatccgtcgacc
Sc9B1.20ΔNrevcyc2ΔN deletion GAGCAGTGCTCCCACGTCCGCCGCGGAGGTGCCGGGCCCtgtaggctggagctgcttc
Sc9B1.20ΔCforwcyc2ΔC deletion GTGCCCTTCCAGAAGGTCGGCCCGTCCGTCATCCCCGACattccggggatccgtcgacc
Sc9B1.20ΔCrevcyc2ΔC deletion CAGTGAACGTCCGTCAGCGCGTCAGTGCGTCAGTGCGGGtgtaggctggagctgcttc
  • The 3′ homology of primers catforw and catrev to the cat gene of pIJ666 and the unique priming sites P1 and P2 of the disruption cassettes are underlined. The oligonucleotides have 39 nt of homology (uppercase) to the relevant gene. Start and stop codons within the 39-nt primer extension are bold.