Table 1.

DNA template and primers used for construction of mutants in this studya

StrainDNA template (reference)Primer 1Primer 2Primer 3Primer 4
toxTΔpro mutantpDH8 (15)5′-CCGGAATTCGAAAATGGTCGATATGAT-3′ (EcoRI)5′-AGTTATCTTAAAATCGCGAATGTGGCTGTTA-3′5′-TAACAGCCACATTCGCGATTTTAAGATAACT-3′5′-CCGGAATTCTACTTTCGAGAAGAACCC-3′ (EcoRI)
tcpAΔpro mutantptcpA::phoA2-1 (36)5′-GCTCTAGACAGACAGATCCACAAGGT-3′ (XbaI)5′-AGCAACACGCACGGTACCGGCCAACTTATTCAATTC-3′ (KpnI)5′-AATAAGTTGGCCGGTACCGTGCGTGTTGCTTACGTT-3′ (KpnI)5′-TCCCCGCGGTGCAATATATGGGAACAT-3′ (SacII)
ΔtcpP mutantO395 chromosomal DNA5′-GGGGTACCGATAACTTTGCAACCGTT-3′ (KpnI)5′-TAATTTTTTGTGCATTACTTTACATTTTCT-3′5′-AGAAAATGTAAAGTAATGCACAAAAAATTA-3′5′-TCCCCGCGGGACGATCTCAATACAACT-3′ (SacII)
ΔtcpH mutantO395 chromosomal DNA5′-GGGGTACCATAAAAAAATGGGTCGTT-3′ (KpnI)5′-ACACTATCTAGGCGGAGCTTTTAATTTTTT-3′5′-AAAAAATTAAAAGCTCCGCCTAGATAGTGT-3′5′-TCCCCGCGGTGTTCTTCTTTTACAAAT-3′ (SacII)
  • a Restriction sites for each individual primer are underlined and in parentheses if applicable. All primers were designed by using the sequences under GenBank accession no.X64098.