TABLE 1.
Oligonucleotide primers
| Primer nameb | Sequence (5′-3′)a | Enzyme | Purpose |
|---|---|---|---|
| EcpleD33-CHis F | GGCATATGACTGCAAAAGTATTAATAG | NdeI | Construction of rEcPleD-CHis expression vector |
| EcpleD33-CHis R | GGCTCGAGTGATAGATATGTGACAACTTTG | XhoI | |
| TRP120 F | CGAGGATCCGATGGATATTGATAATAGTAACATAAG | BamHI | Construction of rTRP120 expression vector |
| TRP120 R | GGTGCGGCCGCTTATACAATATCATTTACTACATTG | NotI | |
| HtrA F | GGACCATGGATCAGAAAGCAGAACAATATG | NcoI | Construction of rHtrA expression vectors |
| HtrAC F | GAACCATGGGCAACGGAATGCCTAAGGATG C | NcoI | Construction of the expression vector for the HtrA C-terminal fragment |
| HtrA R | GGAGTCGACCT CTTCAGCTTGACAGTCA | SalI | Construction of the expression vectors for rHtrA and the HtrA C-terminal fragment |
| Ech0345 F | CCGAAGCTTAATCACTTCGTGGTTGCAAGA | HindIII | Construction of rEch0345 expression vector |
| Ech0345 R | CCGCTCGAGTTATGAAATGTTAGTACTTTTAATTG | XhoI | |
| pleD(I-VMGG) sense | GCAACAGTTTAAAAAACGTGTATCAGACAATATAGTCATGGGCGGCCTATTAGCAAGATTTGGTGGAGAAGAATTTG | Introduction of mutation in PleD I site | |
| pleD(I-VMGG) antisense | CAAATTCTTCTCCACCAAATCTTGCTAATAGGCCGCCCATGACTATATTGTCTGATACACGTTTTTTAAACTGTTGC | ||
| pleD(GG374DE) sense | CGCATTACAGACCTATTAGCAAGATTTGATGAAGAAGAATTTGTTGTAATATTACC | Introduction of mutation in PleD GGDEF motif | |
| pleD(GG374DE) antisense | GGTAATATTACAACAAATTCTTCTTCATCAAATCTTGCTAATAGGTCTGTAATGCG | ||
| adrA real time F | GTCATGTCTGGTGACGTTGC | Detection of adrA mRNA level | |
| adrA real time R | AAGCGGATACAACATAATGACG | ||
| csgD real time F | ACGCTACTGAAGACCAGGAAC | Detection of csgD mRNA level | |
| csgD real time R | GCATTCGCCACGCAGAATAC | ||
| TRP120 gene real time F | GATGACACAGTCTCTCAACC | Detection of TRP120 gene mRNA level | |
| TRP120 gene real time R | CTGATCTGATACACCAGATTC | ||
| ompA real time F | GCAAAAGGCAGAAGAGTATCC | Detection of ompA mRNA level | |
| ompA real time R | GCATTTGCACGCTGTTTG | ||
| p28 real time F | CAGCAGGTAGTGGTATTAACG | Detection of p28 mRNA level | |
| p28 real time R | TCAGTCCAAACACTCCAACTG |
a
Restriction enzyme sites and mutation sites are underlined.
b
F, forward; R, reverse.