TABLE 1.
Oligonucleotides used for PCR amplificationa
| Oligonucleotide | Sequence (5′-3′) | Use |
|---|---|---|
| EpsB1F | GTTAAAGTTGATGATGTTGCC | epsB deletion mutant; left flanking region amplification |
| 1BR | CGGGATCCGTGAGTGAACGTCAATCAC | epsB deletion mutant; left flanking region amplification |
| 2BF | CGGGATCCCCAAAACATTACTAGAAAATC | epsB deletion mutant; right flanking region amplification |
| 2BR | CGTTTCATTTAGATCGGCATTTCCTGAA | epsB deletion mutant; right flanking region amplification |
| EpsC1F | ATAATCTCCAAGACTCTGTCC | epsC deletion mutant; left flanking region amplification |
| EpsC1R | CGGGATCCAGTGTTATCTTG | epsC deletion mutant; left flanking region amplification |
| EpsC2F | CGGGATCCCTTGGAATTGTC | epsC deletion mutant; right flanking region amplification |
| EpsC2R | TAGGTGCTGTTGGTTCAGAG | epsC deletion mutant; right flanking region amplification |
| 1DF | GCTCACATGTCCTCAAACCAAAGCTC | epsD deletion mutant; left flanking region amplification |
| 1DR | CGGGATCCCTCTTCCGTCTTTTTAGCA | epsD deletion mutant; left flanking region amplification |
| 2DF | CGGGATCCGTCGGTTGGAATTAACGCGT | epsD deletion mutant; right flanking region amplification |
| 2DR | CATATTTCGGTTTAATTTCTCTTACGAGAA | epsD deletion mutant; right flanking region amplification |
| 1EF | GCCAGAGTCACCATCTTCACC | epsE deletion mutant; left flanking region amplification |
| 1ER | GCGGATCCCTTTAGCTTGTGACATCTCATTC | epsE deletion mutant; left flanking region amplification |
| 2EF | CGGGATCCGAGTTGGAGCGCGTTAGTACTG | epsE deletion mutant; right flanking region amplification |
| 2ER | CCACTCTAAGTCAATCAAATAACC | epsE deletion mutant; right flanking region amplification |
| EHNF | GCGGATCCATGTCACAAGCTAAAGAGGAAATT | Insertion of epsE open reading frame into pQE-30 to produce His6-EpsE |
| EHNR | CCCAAGCTTCTAACGCGCTCCAACTCTCTTCAAAACAATC | Insertion of epsE open reading frame into pQE-30 to produce His6-EpsE |
| P-eps-F | GGAATTCGCAGACTAGTTTGTAAAAGGACG | eps promoter amplification to construct pJIM4843 |
| P-eps-R | CGCGGATCCCCATTACTCGTATGCTTTTGC | eps promoter amplification to construct pJIM4843 |
a
Oligonucleotide primers were derived from the eps gene sequence of CNRZ1066 (accession number CP000024). In some cases, additional nucleotides were added to create restriction sites (underlined).