TABLE 1.
DNA template and primers used for construction of mutants in this studya
| Strain | DNA template (reference) | Primer 1 | Primer 2 | Primer 3 | Primer 4 |
|---|---|---|---|---|---|
| toxTΔpro mutant | pDH8 (15) | 5′-CCGGAATTCGAAAATGGTCGATATGAT-3′ (EcoRI) | 5′-AGTTATCTTAAAATCGCGAATGTGGCTGTTA-3′ | 5′-TAACAGCCACATTCGCGATTTTAAGATAACT-3′ | 5′-CCGGAATTCTACTTTCGAGAAGAACCC-3′ (EcoRI) |
| tcpAΔpro mutant | ptcpA::phoA2-1 (36) | 5′-GCTCTAGACAGACAGATCCACAAGGT-3′ (XbaI) | 5′-AGCAACACGCACGGTACCGGCCAACTTATTCAATTC-3′ (KpnI) | 5′-AATAAGTTGGCCGGTACCGTGCGTGTTGCTTACGTT-3′ (KpnI) | 5′-TCCCCGCGGTGCAATATATGGGAACAT-3′ (SacII) |
| ΔtcpP mutant | O395 chromosomal DNA | 5′-GGGGTACCGATAACTTTGCAACCGTT-3′ (KpnI) | 5′-TAATTTTTTGTGCATTACTTTACATTTTCT-3′ | 5′-AGAAAATGTAAAGTAATGCACAAAAAATTA-3′ | 5′-TCCCCGCGGGACGATCTCAATACAACT-3′ (SacII) |
| ΔtcpH mutant | O395 chromosomal DNA | 5′-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.