TABLE 2.
Primers used in this study
| Protein | Primer sequences |
|---|---|
| Primers used for construction of expression plasmids | |
| Primers for wild-type alleles | |
| Mlc | TAAGAGGAATAATAAATGTACATGGCTCAA CCCGG, TCCTTCGACCACTTTCATTAA GAG |
| PtsN | CAATTGAGCGAAGTACTTTCATTGGACTG, TTGGTTAACCATGATGTTGTACAGC |
| VC1824 | GCGTGACATCGCGTACATCAG, CATGAATTCCAAGTCACCTTCTTGG |
| Internal primers for construction of mutant alleles | |
| EIIAGlc(H91A) | GCGTTGAGCTGTTTGTTGCCTTCGGTATCG, CGATACCGAAGGCAACAAACAGCTCAACGC |
| EIIAGlc(H91D) | GCGTTGAGCTGTTTGTTGACTTCGGTATCG, CGATACCGAAGTCAACAAACAGCTCAACGC |
| PtsN(H66A) | ATAGCCATCCCAGCCGCACGCATGT, ACATGCGTGCGGCTGGGATGGCTAT |
| PtsN(H66D) | ATAGCCATCCCAGACGCACGCATGT, ACATGCGTGCGTCTGGGATGGCTAT |
| VC1824(H172A) | GGGATTGCTATTCCCGCTGTGATGTTTGC, GCAAACATCACAGCGGGAATAGCAATCCC |
| VC1824(H172D) | GGGATTGCTATTCCCGATGTGATGTTTGC, GCAAACATCACATCGGGAATAGCAATCCC |
| Primers used in quantitative RT-PCR | |
| EI | ACCGTTATCGAAGAGCAAGCCACT, TCTGCGCAGTTTCAGAAGGCGTTA |
| EIIAGlc | GCGATCAAGCCTGCTGGTAACAAA, AAGCCTTCACCTTTCAGCTCAACC |
| HPr | TGTTCAAGCTGCAAACGCTAGGTC, GCAACTAGGTGCTCAACAGCTTCT |
| EIIANtr1 | GCTATTGCAGTGTGAAGAGCCGAT, CTGCGTTACGAAGCTGTTTGAGGA |
| EIIANtr2 | TCGGTATTGATGCTGAACTCGCCT, GGGCTTTGGCATAGTGCCATTTGA |
| ClpX | AGAGTTCATTGGTCGTCTGCCTGT, AACAACGCCGCATACTGTTTGGTC |